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1
Inoue, Takako, Haruyoshi Tomita, and Yasuyoshi Ike. "Bac 32, a Novel Bacteriocin Widely Disseminated among Clinical Isolates of Enterococcus faecium." Antimicrobial Agents and Chemotherapy 50, no. 4 (April 2006): 1202–12. http://dx.doi.org/10.1128/aac.50.4.1202-1212.2006.
Повний текст джерелаАнотація:
ABSTRACT A total of 636 vancomycin-resistant Enterococcus faecium (VRE) isolates that had been obtained between 1994 and 1999 from the Medical School Hospital of the University of Michigan, Ann Arbor, were tested for bacteriocin production. Two hundred seventy-seven (44%) of the strains were bacteriocinogenic; and 193 of these exhibited activity against Enterococcus faecium, Enterococcus hirae, and Enterococcus durans. Strain VRE200 harbors the highly efficient conjugative gentamicin resistance plasmid pG200 (70 kb) and bacteriocin plasmid pTI1 (12.5 kb). The bacteriocin encoded on pTI1 was designated bacteriocin 32 (Bac 32). Bacteriocin 32 was active against E. faecium, E. hirae, and E. durans but showed no activity against Listeria monocytogenes. The Bac 32 genetic locus consists of a bacteriocin gene (bacA) and an immunity gene (bacB). Neither of these genes showed significant homology to any known bacteriocin determinants. The deduced bacA product is 89 amino acids in length, with a putative signal peptide of 19 amino acids at the N terminus. The bacB gene encodes a deduced 55-amino-acid protein without a signal sequence. One hundred eighty-nine strains (97.9%) of the 193 strains with activity against the 3 test enterococcal strains gave rise to the expected specific PCR product with a primer specific for bacA, indicating that there is a high incidence of Bac 32 production among VRE clinical isolates. Data from Southern analyses of plasmid DNA from 189 of the Bac 32-producing strains with a plasmid pTI1-specific probe suggested that 137 (72.5%) of the strains harbored a pTI1-type plasmid. Bac 32 or Bac 32-type bacteriocin activity and the determinant genes were also identified in 22 (39.3%) of a total of 56 vancomycin-sensitive E. faecium clinical isolates, which suggests that this bacteriocin is widely disseminated among E. faecium strains.
2
Malik, Amarila, Elita Yuliantie, Nisa Yulianti Suprahman, Theresa Linardi, Angelina Wening Widiyanti, Jeanita Haldy, Catherine Tjia, and Hiroshi Takagi. "Construction and Functional Analysis of the Recombinant Bacteriocins Weissellicin-MBF from Weissella confusa MBF8-1." Current Pharmaceutical Biotechnology 22, no. 1 (December 31, 2020): 115–22. http://dx.doi.org/10.2174/1389201021666200611111040.
Повний текст джерелаАнотація:
Background: Bacteriocins (Bac1, Bac2, and Bac3) from Weissella confusa MBF8-1, weissellicin- MBF, have been reported as potential alternative substances as well as complements to the existing antibiotics against many antimicrobial-resistant pathogens. Previously, the genes encoded in the large plasmid, pWcMBF8-1, and the spermicidal activity of their synthetic peptides, originally discovered Indonesia, have been studied. Three synthetic bacteriocins peptides of this weissellicin-MBF have been reported for their potential activities, i.e. antibacterial and spermicidal. Objective: The aim of this study was to construct the recombinant Bacteriocin (r-Bac) genes, as well as to investigate the gene expressions and their functional analysis. Method: Here, the recombinant Bacteriocin (r-Bac) genes were constructed and the recombinant peptides (r-Bac1, r-Bac2, and r-Bac3) in B. subtilis DB403 cells were produced on a large scale. After purification, using the His-tag affinity column, their potential bioactivities were measured as well as their antibacterial minimum inhibitory concentrations against Leuconostoc mesenteroides and Micrococcus luteus, were determined. Results: Pure His-tag-recombinant Bac1, Bac2, and Bac3 were obtained and they could inhibit the growth of L. mesenteroides and M. luteus. Conclusion: The recombinant bacteriocin could be obtained although with weak activity in inhibiting gram-positive bacterial growth.
3
Murrell, Isa, Gavin S. Wilkie, Andrew J. Davison, Evelina Statkute, Ceri A. Fielding, Peter Tomasec, Gavin W. G. Wilkinson, and Richard J. Stanton. "Genetic Stability of Bacterial Artificial Chromosome-Derived Human Cytomegalovirus during CultureIn Vitro." Journal of Virology 90, no. 8 (February 3, 2016): 3929–43. http://dx.doi.org/10.1128/jvi.02858-15.
Повний текст джерелаАнотація:
ABSTRACTClinical human cytomegalovirus (HCMV) strains invariably mutate when propagatedin vitro. Mutations in gene RL13 are selected in all cell types, whereas in fibroblasts mutants in the UL128 locus (UL128L; genes UL128, UL130, and UL131A) are also selected. In addition, sporadic mutations are selected elsewhere in the genome in all cell types. We sought to investigate conditions under which HCMV can be propagated without incurring genetic defects. Bacterial artificial chromosomes (BACs) provide a stable, genetically defined source of viral genome. Viruses were generated from BACs containing the genomes of strains TR, TB40, FIX, and Merlin, as well as from Merlin-BAC recombinants containing variant nucleotides in UL128L from TB40-BAC4 or FIX-BAC. Propagation of viruses derived from TR-BAC, TB40-BAC4, and FIX-BAC in either fibroblast or epithelial cells was associated with the generation of defects around the prokaryotic vector, which is retained in the unique short (US) region of viruses. This was not observed for Merlin-BAC, from which the vector is excised in derived viruses; however, propagation in epithelial cells was consistently associated with mutations in the unique longb′ (UL/b′) region, all impacting on gene UL141. Viruses derived from Merlin-BAC in fibroblasts had mutations in UL128L, but mutations occurred less frequently with recombinants containing UL128L nucleotides from TB40-BAC4 or FIX-BAC. Viruses derived from a Merlin-BAC derivative in which RL13 and UL128L were either mutated or repressed were remarkably stable in fibroblasts. Thus, HCMV containing a wild-type gene complement can be generatedin vitroby deriving virus from a self-excising BAC in fibroblasts and repressing RL13 and UL128L.IMPORTANCEResearchers should aim to study viruses that accurately represent the causative agents of disease. This is problematic for HCMV because clinical strains mutate rapidly when propagatedin vitro, becoming less cell associated, altered in tropism, more susceptible to natural killer cells, and less pathogenic. Following isolation from clinical material, HCMV genomes can be stabilized by cloning into bacterial artificial chromosomes (BACs), and then virus is regenerated by DNA transfection. However, mutations can occur not only during isolation prior to BAC cloning but also when virus is regenerated. We have identified conditions under which BAC-derived viruses containing an intact, wild-type genome can be propagatedin vitrowith minimal risk of mutants being selected, enabling studies of viruses expressing the gene complement of a clinical strain. However, even under these optimized conditions, sporadic mutations can occur, highlighting the advisability of sequencing the HCMV stocks used in experiments.
4
Yamashita, Hitoshi, Haruyoshi Tomita, Takako Inoue, and Yasuyoshi Ike. "Genetic Organization and Mode of Action of a Novel Bacteriocin, Bacteriocin 51: Determinant of VanA-Type Vancomycin-Resistant Enterococcus faecium." Antimicrobial Agents and Chemotherapy 55, no. 9 (June 27, 2011): 4352–60. http://dx.doi.org/10.1128/aac.01274-10.
Повний текст джерелаАнотація:
ABSTRACTBacteriocin 51 (Bac 51) is encoded on the mobile plasmid pHY (6,037 bp), which was isolated from vancomycin-resistantEnterococcus faeciumVRE38. Bacteriocin 51 is active againstE. faecium,E. hirae, andE. durans. Sequence analysis of pHY showed that it encodes nine open reading frames (ORFs) from ORF1 to ORF9 (in that order). Genetic analysis suggested that ORF1 and ORF2, which were designatedbacAandbacB, respectively, are the bacteriocin and immunity genes.bacAencodes a 144-amino-acid protein. The deduced BacA protein has a typical signal sequence at its amino terminus, and a potential signal peptidase-processing site corresponding to the V-E-A sequence is located between the 37th and 39th amino acids. The predicted mature BacA protein consists of 105 amino acids. A potential promoter sequence was identified upstream of the start codon.bacBencodes a 55-amino-acid protein. No obvious promoter or terminator sequence was identified betweenbacAandbacB. Northern blot analysis ofbacAandbacBwith abacARNA probe produced a transcript of approximately 700 nucleotides, which corresponded to the combined nucleotide sizes ofbacAandbacB, indicating that transcription was initiated from the promoter upstream ofbacA, continued throughbacB, and was terminated at the terminator downstream ofbacB. The transcription start site was determined to be the T nucleotide located 6 nucleotides downstream from the −10 promoter sequence. These results indicate thatbacAandbacBconstitute an operon and thatbacAis the bacteriocin structural gene whilebacBis the immunity gene. The purified C-terminally His tagged BacA protein of Bac 51 showed bacteriostatic activity against the indicator strain. The purified C-terminally His tagged BacA protein of Bac 32 (whose mature BacA protein has 54 amino acids) and the culture filtrates of the Bac 31- and Bac 43-producingE. faecalisstrain FA2-2 showed bactericidal activity. Bac 31 and Bac 43 are pore-forming bacteriocins, unlike the newly characterized bacteriocin Bac 51.
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Oh, Seungdae, Zohre Kurt, Despina Tsementzi, Michael R. Weigand, Minjae Kim, Janet K. Hatt, Madan Tandukar, Spyros G. Pavlostathis, Jim C. Spain, and Konstantinos T. Konstantinidis. "Microbial Community Degradation of Widely Used Quaternary Ammonium Disinfectants." Applied and Environmental Microbiology 80, no. 19 (June 20, 2014): 5892–900. http://dx.doi.org/10.1128/aem.01255-14.
Повний текст джерелаАнотація:
ABSTRACTBenzalkonium chlorides (BACs) are disinfectants widely used in a variety of clinical and environmental settings to prevent microbial infections, and they are frequently detected in nontarget environments, such as aquatic and engineered biological systems, even at toxic levels. Therefore, microbial degradation of BACs has important ramifications for alleviating disinfectant toxicity in nontarget environments as well as compromising disinfectant efficacy in target environments. However, how natural microbial communities respond to BAC exposure and what genes underlie BAC biodegradation remain elusive. Our previous metagenomic analysis of a river sediment microbial community revealed that BAC exposure selected for a low-diversity community, dominated by several members of thePseudomonasgenus that quickly degraded BACs. To elucidate the genetic determinants of BAC degradation, we conducted time-series metatranscriptomic analysis of this microbial community during a complete feeding cycle with BACs as the sole carbon and energy source under aerobic conditions. Metatranscriptomic profiles revealed a candidate gene for BAC dealkylation, the first step in BAC biodegradation that results in a product 500 times less toxic. Subsequent biochemical assays and isolate characterization verified that the putative amine oxidase gene product was functionally capable of initiating BAC degradation. Our analysis also revealed cooperative interactions among community members to alleviate BAC toxicity, such as the further degradation of BAC dealkylation by-products by organisms not encoding amine oxidase. Collectively, our results advance the understanding of BAC aerobic biodegradation and provide genetic biomarkers to assess the critical first step of this process in nontarget environments.
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Shah, Raina J., Charles G. Burhans, Michael J. Wise, J. Paul Frantz, and Timothy P. Rhoades. "Exploring the relationship between risk perception and U.S. driver acceptance of a 0.05% BAC limit." Proceedings of the Human Factors and Ergonomics Society Annual Meeting 60, no. 1 (September 2016): 1661–65. http://dx.doi.org/10.1177/1541931213601382.
Повний текст джерелаАнотація:
The National Transportation Safety Board (NTSB) has recently recommended that states reduce the legal blood alcohol concentration (BAC) limit for driving from 0.08% to 0.05%. This study assessed attitudes of drivers toward this recommendation. It also assessed the extent to which attitudes toward lowering the legal BAC limit were related to the perceived risk of driving at different BAC levels as compared to other factors. A majority of drivers surveyed (54%) were in favor of the 0.05% BAC limit proposal, with 26% against it and 20% neutral toward it. Those who believed that driving at 0.05% BAC posed an increased risk for a crash were more likely to support the proposal than those who did not think that driving at 0.05% BAC posed an increased crash risk. However, perceptions about the risk posed by driving at various BACs did not completely determine policy preferences. Some participants supported the 0.05% BAC limit even though they did not perceive a risk reduction benefit, while others perceived a risk reduction benefit of a 0.05% BAC limit but still did not support the policy. Implications for the role of risk perceptions in policy preferences are discussed.
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Sinzger, Christian, Gabriele Hahn, Margarete Digel, Ruth Katona, Kerstin Laib Sampaio, Martin Messerle, Hartmut Hengel, Ulrich Koszinowski, Wolfram Brune, and Barbara Adler. "Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E." Journal of General Virology 89, no. 2 (February 1, 2008): 359–68. http://dx.doi.org/10.1099/vir.0.83286-0.
Повний текст джерелаАнотація:
Human cytomegalovirus (HCMV) strain TB40/E, replicates efficiently, exhibits a broad cell tropism and is widely used for infection of endothelial cells and monocyte-derived cells yet has not been available in a phenotypically homogeneous form compatible with genetic analysis. To overcome this problem, we cloned the TB40/E strain into a bacterial artificial chromosome (BAC) vector. Both highly endotheliotropic and poorly endotheliotropic virus clones, representing three distinct restriction fragment patterns, were reconstituted after transfection of BAC clones derived from previously plaque-purified strain TB40/E. For one of the highly endotheliotropic clones, TB40-BAC4, we provide the genome sequence. Two BACs with identical restriction fragment patterns but different cell tropism were further analysed in the UL128-UL131A gene region. Sequence analysis revealed one coding-relevant adenine insertion at position 332 of UL128 in the BAC of the poorly endotheliotropic virus, which caused a frameshift in the C-terminal part of the coding sequence. Removal of this insertion by markerless mutagenesis restored the highly endotheliotropic phenotype, indicating that the loss of endothelial cell tropism was caused by this insertion. In conclusion, HCMV strain TB40/E, which combines the high endothelial cell tropism of a clinical isolate with the high titre growth of a cell culture adapted strain, is now available as a BAC clone suitable for genetic engineering. The results also suggest BAC cloning as a suitable method for selection of genetically defined virus clones.
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Schumacher, Daniel, B. Karsten Tischer, Walter Fuchs, and Nikolaus Osterrieder. "Reconstitution of Marek's Disease Virus Serotype 1 (MDV-1) from DNA Cloned as a Bacterial Artificial Chromosome and Characterization of a Glycoprotein B-Negative MDV-1 Mutant." Journal of Virology 74, no. 23 (December 1, 2000): 11088–98. http://dx.doi.org/10.1128/jvi.74.23.11088-11098.2000.
Повний текст джерелаАнотація:
ABSTRACT The complete genome of Marek's disease virus serotype 1 (MDV-1) strain 584Ap80C was cloned in Escherichia coli as a bacterial artificial chromosome (BAC). BAC vector sequences were introduced into the US2 locus of the MDV-1 genome by homologous recombination. Viral DNA containing the BAC vector was used to transform Escherichia coli strain DH10B, and several colonies harboring the complete MDV-1 genome as an F plasmid (MDV-1 BACs) were identified. DNA from various MDV-1 BACs was transfected into chicken embryo fibroblasts, and from 3 days after transfection, infectious MDV-1 was obtained. Growth of MDV-1 recovered from BACs was indistinguishable from that of the parental virus, as assessed by plaque formation and determination of growth curves. In one of the MDV-1 BAC clones, sequences encoding glycoprotein B (gB) were deleted by one-step mutagenesis using a linear DNA fragment amplified by PCR. Mutant MDV-1 recovered after transfection of BAC DNA that harbored a 2.0-kbp deletion of the 2.6-kbp gB gene were able to grow and induce MDV-1-specific plaques only on cells providing MDV-1 gB in trans. The gB-negative virus reported here represents the first MDV-1 mutant with a deletion of an essential gene and demonstrates the power and usefulness of BACs to analyze genes and gene products in slowly growing and strictly cell-associated herpesviruses.
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Zhao, S. "A comprehensive BAC resource." Nucleic Acids Research 29, no. 1 (January 1, 2001): 141–43. http://dx.doi.org/10.1093/nar/29.1.141.
Повний текст джерела
10
Wang, Min Qi, Mary E. Nicholson, Beverly S. Mahoney, Yuhua Li, and Mike A. Perko. "Proprioceptive Responses under Rising and Falling BACs: A Test of the Mellanby Effect." Perceptual and Motor Skills 77, no. 1 (August 1993): 83–88. http://dx.doi.org/10.2466/pms.1993.77.1.83.
Повний текст джерелаАнотація:
This study examined proprioceptive responses under equivalent rising and falling blood alcohol concentrations (BAC), using a repeated-measures design. Seven volunteer subjects, 21 to 35 years of age, participated in the study. After alcohol consumption, BAC readings were obtained every 5 minutes, and the proprioceptive responses were measured at the following BAC levels (in %): 0 (baseline), rising 0.05, 0.075, 0.1, falling 0.075, and 0.05. The analysis focused on the comparisons of these measures at the equivalent rising and falling 0.05% and at the 0.075% BACs. Results showed that the proprioceptive response was less accurate during the rising than the falling BACs.
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Moi, John H. Y., Uyen Phan, Adam de Gruchy, Danny Liew, Tanya I. Yuen, John E. Cunningham, and Ian P. Wicks. "Is establishing a specialist back pain assessment and management service in primary care a safe and effective model? Twelve-month results from the Back pain Assessment Clinic (BAC) prospective cohort pilot study." BMJ Open 8, no. 10 (October 2018): e019275. http://dx.doi.org/10.1136/bmjopen-2017-019275.
Повний текст джерелаАнотація:
ObjectivesTo report on the design, implementation and evaluation of the safety and effectiveness of the Back pain Assessment Clinic (BAC) model.DesignBAC is a new, community-based specialist service for assessing and managing neck and low back pain (LBP). The BAC pilot was supported by a Victorian Department of Health and Human Services grant and was evaluated using the Victorian Innovation Reform Impact Assessment Framework (VIRIAF). Data were obtained by auditing BAC activity (22 July 2014 to 30 June 2015) and conducting surveys and interviews of patients, stakeholders and referrers.SettingTertiary and primary care.ParticipantsAdult patients with neck and LBP referred for outpatient surgical consultation.Main outcome measuresVIRIAF outcomes: (1) access to care; (2) appropriate and safe care; (3) workforce optimisation and integration; and (4) efficiency and sustainability.ResultsA total of 522 patients were seen during the pilot. Most were referred to hospital services by general practitioners (87%) for LBP (63%) and neck pain (24%). All patients were seen within 10 weeks of referral and commenced community-based allied health intervention within 2–4 weeks of assessment in BAC. Of patients seen, 34% had medications adjusted, 57% were referred for physiotherapy, 3.2% to pain services, 1.1% to rheumatology and 1.8% for surgical review. Less MRI scans were ordered in BAC (6.4%) compared with traditional spinal surgical clinics (89.8%), which translated to a cost-saving of $52 560 over 12 months. Patient and staff satisfaction was high. There have been no patient complaints or adverse incidents.ConclusionEvaluation of the BAC pilot suggests it is a potentially safe and cost-saving alternative model of care. Results of the BAC pilot merit further evaluation to determine the potential cost-effectiveness, longer term and broader societal impact of implementing BAC more widely.
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Cao, Yicheng, Hyung Lyun Kang, Xuequn Xu, Mei Wang, So Hee Dho, Jun Ryul Huh, Byeong-Jae Lee, et al. "A 12-Mb Complete Coverage BAC Contig Map in Human Chromosome 16p13.1–p11.2." Genome Research 9, no. 8 (August 1, 1999): 763–74. http://dx.doi.org/10.1101/gr.9.8.763.
Повний текст джерелаАнотація:
We have constructed a complete coverage BAC contig map that spans a 12-Mb genomic segment in the human chromosome 16p13.1–p11.2 region. The map consists of 68 previously mapped STSs and 289 BAC clones, 51 of which—corresponding to a total of 7.721 Mb of genomic DNA—have been sequenced, and provides a high resolution physical map of the region. Contigs were initially built based mainly on the analysis of STS contents and restriction fingerprint patterns of the clones. To close the gaps, probes derived from BAC clone ends were used to screen deeper BAC libraries. Clone end sequence data obtained from chromosome 16-specific BACs, as well as from public databases, were used for the identification of BACs that overlap with fully sequenced BACs by means of sequence match. This approach allowed precise alignment of clone overlaps in addition to restriction fingerprint comparison. A freehand contig drawing software tool was developed and used to manage the map data graphically and generate a real scale physical map. The map we present here is ∼3.5 × deep and provides a minimal tiling path that covers the region in an array of contigous, overlapping BACs.
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Zhou, Fuchun, Qiuhua Li, Scott W. Wong, and Shou-Jiang Gao. "Autoexcision of Bacterial Artificial Chromosome Facilitated by Terminal Repeat-Mediated Homologous Recombination: a Novel Approach for Generating Traceless Genetic Mutants of Herpesviruses." Journal of Virology 84, no. 6 (January 13, 2010): 2871–80. http://dx.doi.org/10.1128/jvi.01734-09.
Повний текст джерелаАнотація:
ABSTRACT Infectious bacterial artificial chromosomes (BACs) of herpesviruses are powerful tools for genetic manipulation. However, the presence of BAC vector sequence in the viral genomes often causes genetic and phenotypic alterations. While the excision of the BAC vector cassette can be achieved by homologous recombination between extra duplicate viral sequences or loxP site-mediated recombination, these methods either are inefficient or leave a loxP site mark in the viral genome. Here we describe the use of viral intrinsic repeat sequences, which are commonly present in herpesviral genomes, to excise the BAC vector cassette. Using a newly developed in vitro transposon-based cloning approach, we obtained an infectious BAC of rhesus rhadinovirus (RRV) strain RRV26-95 with the BAC vector cassette inserted in the terminal repeat (TR) region. We showed that the BAC vector cassette was rapidly excised upon reconstitution in cells predominantly through TR-mediated homologous recombination. Genetic and phenotypic analysis showed that the BAC-excised virus was reversed to wild-type RRV. Using this autoexcisable BAC clone, we successfully generated an RRV mutant with a deletion of Orf50, which encodes a replication and transcription activator (RTA) protein. Together, these results illustrate the usefulness of TR for genetic manipulation of herpesviruses when combined with the novel transposon-based cloning approach.
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Redwood, Alec J., Martin Messerle, Nicole L. Harvey, Christopher M. Hardy, Ulrich H. Koszinowski, Malcolm A. Lawson, and Geoffrey R. Shellam. "Use of a Murine Cytomegalovirus K181-Derived Bacterial Artificial Chromosome as a Vaccine Vector for Immunocontraception." Journal of Virology 79, no. 5 (March 1, 2005): 2998–3008. http://dx.doi.org/10.1128/jvi.79.5.2998-3008.2005.
Повний текст джерелаАнотація:
ABSTRACT Cytomegaloviruses (CMVs) are members of the Betaherpesvirinae subfamily of the Herpesviridae, and their properties of latency, large DNA size, gene redundancy, and ability to be cloned as bacterial artificial chromosomes (BACs) suggest their utility as vaccine vectors. While the K181 strain of murine CMV (MCMV) is widely used to study MCMV biology, a BAC clone of this virus had not previously been produced. We report here the construction of a BAC clone of the K181Perth strain of MCMV. The in vivo and in vitro growth characteristics of virus derived from the K181 BAC were similar to those of wild-type K181. The utility of the K181 BAC as a method for the rapid production of vaccine vectors was assessed. A vaccine strain of BAC virus, expressing the self-fertility antigen, murine zona pellucida 3, was produced rapidly using standard bacterial genetics techniques and rendered female BALB/c mice infertile with a single intraperitoneal inoculation. In addition, attenuated vaccine strains lacking the open reading frames m07 to m12 exhibited no reduction in efficacy compared to the full-length vaccine strain. In conclusion, we describe the production of a K181-based BAC virus which behaved essentially as wild-type K181 and allowed the rapid production of effective viral vaccine vectors.
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Lewers, K. S., S. D. Nilmalgoda, A. L. Warner, H. T. Knap, and B. F. Matthews. "Physical mapping of resistant and susceptible soybean genomes near the soybean cyst nematode resistance gene Rhg4." Genome 44, no. 6 (December 1, 2001): 1057–64. http://dx.doi.org/10.1139/g01-109.
Повний текст джерелаАнотація:
The soybean cyst nematode (SCN), Heterodera glycines Ichinohe, is the foremost pest of soybean (Glycine max L. Merr.). The rhg1 allele on linkage group (LG) G and the Rhg4 allele on LG A2 are important in conditioning resistance. Markers closely linked to the Rhg4 locus were used previously to screen a library of bacterial artificial chromosome (BAC) clones from susceptible 'Williams 82' and identified a single 150-kb BAC, Gm_ISb001_056_G02 (56G2). End-sequenced subclones positioned onto a restriction map provided landmarks for identifying the corresponding region from a BAC library from accession PI 437654 with broad resistance to SCN. Seventy-three PI 437654 BACs were assigned to contigs based upon HindIII restriction fragment profiles. Four contigs represented the PI 437654 counterpart of the 'Williams 82' BAC, with PCR assays connecting these contigs. Some of the markers on the PI 437654 contigs are separated by a greater physical distance than in the 'Williams 82' BAC and some primers amplify bands from BACs in the mid-portion of the connected PI 437654 BAC contigs that are not amplified from the 'Williams 82' BAC. These observations suggest that there is an insertion in the PI 437654 genome relative to the 'Williams 82' genome in the Rhg4 region.Key words: BAC, deletion, insertion, resistance gene, soybean cyst nematode.
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Van de Wiel, Albert, David Moolenaar, and Jos Wielders. "The Bac(chus) experiment: blood alcohol concentrations after wine tasting." Wine Studies 2, no. 1 (March 8, 2012): 1. http://dx.doi.org/10.4081/ws.2012.e1.
Повний текст джерелаАнотація:
Blood alcohol concentrations (BACs) were measured in ten volunteers after a wine tasting event with and without the swallowing of 15 mL of each wine. In case ten wines were tasted within one hour without swallowing, buccal mucosa absorption did not result in problematic BAC’s; however in case 15 mL of each wine was swallowed, BAC’s may exceed the legal driving limit of most countries. It is recommended to eat beforehand, but also to wait at least one hour after the session before driving back home.
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Marra, Marco A., Martin Krzywinski, Readman Chiu, Matthew Field, Inanc Birol, Brian D’Souza, Ian Bosdet, et al. "Towards the Human Cancer Genome Project: A Sequence-Ready Physical Map of a Follicular Lymphoma Genome." Blood 106, no. 11 (November 16, 2005): 605. http://dx.doi.org/10.1182/blood.v106.11.605.605.
Повний текст джерелаАнотація:
Abstract With the aim of identifying and sequencing mutations in follicular lymphoma genomes, we have begun a project to generate at least 24 deeply redundant sequence-ready Bacterial Artificial Clone (BAC) - based whole genome maps, each from a different individual’s lymphoma. BAC-array CGH and Affymetrix whole-genome sampling assays (WGSA) will be used along with the mapping data to identify genomic amplifications and losses in the lymphomas. Results from the mapping and array studies will be used to prioritize BAC clones for sequence analysis. Because each map will span essentially the entire genome of the corresponding lymphoma, we anticipate that essentially all regions of each tumor genome will be represented in easily sequenced BAC clones. This approach facilitates targeted sequencing of genomic regions of interest, including those containing genes relevant to cancer or harboring amplifications or deletions. Our mapping strategy hinges on the successful creation of deeply redundant high quality BAC libraries from primary lymphomas and large scale high throughput restriction enzyme fingerprinting of individual BACs with a version of the technology we used to map the human, mouse, rat and other genomes. The effort is large-scale, and will result in the generation of at least 2.5 million fingerprinted BAC clones over the next three years. Using the fingerprints, we will align the BACs to the reference human genome to assess genome coverage and to identify candidate genome rearrangements. In parallel, we will assemble the fingerprints into genome maps, looking for larger-scale genome variations between the lymphoma maps and the reference genome sequence. To test the feasibility of our approach, we obtained two restriction digest fingerprints from each of 140,000 individual BAC clones. BACs were sampled from a 7-fold redundant BAC library that had been created from genomic DNA purified from a primary follicular lymphoma sample. The fingerprints are being assembled into a clone map with the intent of reconstructing the entire tumor genome. 90,377 fingerprinted clones with unambiguous single alignments to the reference sequence were automatically assembled into 15,538 contigs. Subsequent rounds of semi-automatic contig merging further reduced the number of contigs to 5,433. Only 1,241 clones remained unassembled. We anchored the tumor genome map to the reference human genome sequence by aligning the clone fingerprints to the restriction map computed from the reference sequence assembly. As a result of this, we identified a BAC that captured the canonical t(14;18) translocation characteristic of follicular lymphomas. We sequenced this BAC and confirmed that it contains the expected translocation. Almost 2.6 gigabases (~91%) of the reference genome are represented in the evolving map, with an additional 50,000 clone fingerprints awaiting incorporation into the map assembly. Among these are repeat-rich and other clones that may well harbor genome rearrangements. Additional prioritization of sequencing targets will be undertaken when map construction and analysis of genome copy number alterations are complete.
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Kubát, Z. "Chromosome walking with BAC clones as a method of genome mapping." Plant, Soil and Environment 53, No. 10 (January 7, 2008): 447–50. http://dx.doi.org/10.17221/2198-pse.
Повний текст джерелаАнотація:
Current sequencing projects are often based on random sequencing of genomic libraries followed by contig assembly by means of bioinformatics tools. This approach is convenient for whole genome sequencing projects. Chromosome walking described here is suitable for mapping and sequencing of short genomic regions in species where whole genome sequencing is not possible or for cloning gene from its closest known marker. This method is based on searching for overlapping BAC clones specific for the genomic region of interest.
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Kuhl, Heiner, Elena Sarropoulou, Mbaye Tine, Georgios Kotoulas, Antonios Magoulas, and Richard Reinhardt. "A Comparative BAC Map for the Gilthead Sea Bream (Sparus aurataL.)." Journal of Biomedicine and Biotechnology 2011 (2011): 1–7. http://dx.doi.org/10.1155/2011/329025.
Повний текст джерелаАнотація:
This study presents the first comparative BAC map of the gilthead sea bream (Sparus aurata), a highly valuated marine aquaculture fish species in the Mediterranean. High-throughput end sequencing of a BAC library yielded 92,468 reads (60.6 Mbp). Comparative mapping was achieved by anchoring BAC end sequences to the three-spined stickleback (Gasterosteus aculeatus) genome. BACs that were consistently ordered along the stickleback chromosomes accounted for 14,265 clones. A fraction of 5,249 BACs constituted a minimal tiling path that covers 73.5% of the stickleback chromosomes and 70.2% of the genes that have been annotated. The N50 size of 1,485 “BACtigs” consisting of redundant BACs is 337,253 bp. The largest BACtig covers 2.15 Mbp in the stickleback genome. According to the insert size distribution of mapped BACs the sea bream genome is 1.71-fold larger than the stickleback genome. These results represent a valuable tool to researchers in the field and may support future projects to elucidate the whole sea bream genome.
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Armas, Federica, Adriana Di Stasi, Mario Mardirossian, Antonello A. Romani, Monica Benincasa, and Marco Scocchi. "Effects of Lipidation on a Proline-Rich Antibacterial Peptide." International Journal of Molecular Sciences 22, no. 15 (July 26, 2021): 7959. http://dx.doi.org/10.3390/ijms22157959.
Повний текст джерелаАнотація:
The emergence of multidrug-resistant bacteria is a worldwide health problem. Antimicrobial peptides have been recognized as potential alternatives to conventional antibiotics, but still require optimization. The proline-rich antimicrobial peptide Bac7(1-16) is active against only a limited number of Gram-negative bacteria. It kills bacteria by inhibiting protein synthesis after its internalization, which is mainly supported by the bacterial transporter SbmA. In this study, we tested two different lipidated forms of Bac7(1-16) with the aim of extending its activity against those bacterial species that lack SbmA. We linked a C12-alkyl chain or an ultrashort cationic lipopeptide Lp-I to the C-terminus of Bac7(1-16). Both the lipidated Bac-C12 and Bac-Lp-I forms acquired activity at low micromolar MIC values against several Gram-positive and Gram-negative bacteria. Moreover, unlike Bac7(1-16), Bac-C12, and Bac-Lp-I did not select resistant mutants in E. coli after 14 times of exposure to sub-MIC concentrations of the respective peptide. We demonstrated that the extended spectrum of activity and absence of de novo resistance are likely related to the acquired capability of the peptides to permeabilize cell membranes. These results indicate that C-terminal lipidation of a short proline-rich peptide profoundly alters its function and mode of action and provides useful insights into the design of novel broad-spectrum antibacterial agents.
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Musile, Giacomo, Nicola Pigaiani, Daniela Sorio, Michela Colombari, Federica Bortolotti, and Franco Tagliaro. "Alcohol-associated traffic injuries in Verona territory: A nine-year survey." Medicine, Science and the Law 61, no. 1_suppl (January 2021): 7–13. http://dx.doi.org/10.1177/0025802420937577.
Повний текст джерелаАнотація:
According to the World Health Organization, as many as 25% of traffic accidents are linked to alcohol abuse. This study describes the results of a nine-year study performed on injured drivers ( N = 12,806) in the Verona area of Northern Italy. Blood samples were mandatorily collected on injured drivers who were admitted to the Emergency Health Care Unit of Verona Hospital between 2009 and 2017, after they had been involved in a traffic accident. Blood alcohol concentration (BAC) determination was then undertaken using a validated head space–gas chromatography–flame ionisation detector (HS-GC-FID) method. We found that 21% of drivers tested positive for alcohol (BAC ≥0.01 g/L), while 16.8% presented with BAC levels above the Italian legal limit (>0.5 g/L). Of those who had positive BACs, about 50% presented with very high BAC levels (>1.5 g/L). Daily time distribution analyses, involving 2031 alcohol-positive drivers, showed a surge between 18:00 hours and 06:00 hours (74.3%), with a specific rise during the weekend (58.9%). The percentage of alcohol-related road accidents was 20.6%, which is lower than results reported in other international studies performed over the last 20 years. However, evidence that around 50% of the positive subjects showed a BAC >1.5 g/L confirms the correlation between BAC and accident risk, which becomes even more significant at progressively increasing levels of BAC. The study highlights the need to implement further strategies to both prevent and deter the use of alcohol while driving.
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Martins, Lívia do Vale, Ana Rafaela da Silva Oliveira, Maria Muñoz Amatriaín, Timothy J. Close, Andrea Pedrosa Harand, Lidiane de Lima Feitoza, and Ana Christina Brasileiro Vidal. "Comparative cytogenetic analysis in Vigna sp. revealed by BAC-FISH." Semina: Ciências Biológicas e da Saúde 38, no. 1supl (February 16, 2018): 138. http://dx.doi.org/10.5433/1679-0367.2017v38n1suplp138.
Повний текст джерелаАнотація:
Vigna Savi species constitute a group of worldwide legumes of great socioeconomic importance. Progress in its genomic resources allowed the development of a consensus genetic map with a high-density of SNP markers and construction of Bacterial Artificial Chromosome (BAC) libraries. Fluorescent in situ hybridization (FISH) using BAC clones as probes (BAC-FISH) became a powerful tool for synteny and collinearity analyses among closely related species in several groups of plants. Previous work using BAC-FISH with probes of the P. vulgaris ‘BAT93’ library of V. unguiculata (Vu) and V. aconitifolia (Vac) chromosomes showed partial conservation of macrosynteny with chromosomal rearrangements. For a better understanding of the genomic organization and karyotype evolution in Vigna, we performed a comparative cytogenetic study with three Vigna species (2n = 22), using BAC-FISH. Four single-copy BACs of chromosome 3 and chromosome 11 of V. unguiculata (Vu3 and Vu11, respectively) were hybridized in situ on mitotic metaphase chromosomes of V. angularis (Van) and V. radiata (Vr). For chromosome 3, two Vu3 BACs (H31G07 and H50P11) were located in the same orientation at Vu, Van and Vr chromosomes, in interstitial regions in the short and long arms, respectively. These results suggest a conservation of synteny for both markers. On the other hand, H49E24 and H85I15 BACs were located respectively in terminal region in the short arm and subterminal region in the long arm of Vu11, while a distinct pattern was observed in Van, which showed both BACs at adjacent positions in the short arm. This suggests that a pericentric inversion involving BAC H85I15 occurred in one of these species. Based on the present results, associated with our previous works, the inversion observed on Vu11 seems to have occurred in V. unguiculata after Vigna and Phaseolus separation, since position of other BAC clones were different in Vu when compared to Vac, Van and Pv. These data demonstrate the feasibility of the BAC-FISH technique in comparative chromosome mapping, showing a break of macrosynteny among species of Phaseoloid clade. This is an initial step of an extensive macrosynteny study with BAC-FISH in Vigna.
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Brosch, Roland, Stephen V. Gordon, Alain Billault, Thierry Garnier, Karin Eiglmeier, Catherine Soravito, Bart G. Barrell, and Stewart T. Cole. "Use of a Mycobacterium tuberculosisH37Rv Bacterial Artificial Chromosome Library for Genome Mapping, Sequencing, and Comparative Genomics." Infection and Immunity 66, no. 5 (May 1, 1998): 2221–29. http://dx.doi.org/10.1128/iai.66.5.2221-2229.1998.
Повний текст джерелаАнотація:
ABSTRACT The bacterial artificial chromosome (BAC) cloning system is capable of stably propagating large, complex DNA inserts in Escherichia coli. As part of the Mycobacterium tuberculosis H37Rv genome sequencing project, a BAC library was constructed in the pBeloBAC11 vector and used for genome mapping, confirmation of sequence assembly, and sequencing. The library contains about 5,000 BAC clones, with inserts ranging in size from 25 to 104 kb, representing theoretically a 70-fold coverage of the M. tuberculosisgenome (4.4 Mb). A total of 840 sequences from the T7 and SP6 termini of 420 BACs were determined and compared to those of a partial genomic database. These sequences showed excellent correlation between the estimated sizes and positions of the BAC clones and the sizes and positions of previously sequenced cosmids and the resulting contigs. Many BAC clones represent linking clones between sequenced cosmids, allowing full coverage of the H37Rv chromosome, and they are now being shotgun sequenced in the framework of the H37Rv sequencing project. Also, no chimeric, deleted, or rearranged BAC clones were detected, which was of major importance for the correct mapping and assembly of the H37Rv sequence. The minimal overlapping set contains 68 unique BAC clones and spans the whole H37Rv chromosome with the exception of a single gap of ∼150 kb. As a postgenomic application, the canonical BAC set was used in a comparative study to reveal chromosomal polymorphisms between M. tuberculosis, M. bovis, and M. bovis BCG Pasteur, and a novel 12.7-kb segment present in M. tuberculosis but absent from M. bovis and M. bovis BCG was characterized. This region contains a set of genes whose products show low similarity to proteins involved in polysaccharide biosynthesis. The H37Rv BAC library therefore provides us with a powerful tool both for the generation and confirmation of sequence data as well as for comparative genomics and other postgenomic applications. It represents a major resource for present and future M. tuberculosis research projects.
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Rondelet, Arnaud, Andrei Pozniakovsky, Devika Namboodiri, Richard Cardoso da Silva, Divya Singh, Marit Leuschner, Ina Poser, et al. "ESI mutagenesis: a one-step method for introducing mutations into bacterial artificial chromosomes." Life Science Alliance 4, no. 2 (December 8, 2020): e202000836. http://dx.doi.org/10.26508/lsa.202000836.
Повний текст джерелаАнотація:
Bacterial artificial chromosome (BAC)–based transgenes have emerged as a powerful tool for controlled and conditional interrogation of protein function in higher eukaryotes. Although homologous recombination-based recombineering methods have streamlined the efficient integration of protein tags onto BAC transgenes, generating precise point mutations has remained less efficient and time-consuming. Here, we present a simplified method for inserting point mutations into BAC transgenes requiring a single recombineering step followed by antibiotic selection. This technique, which we call exogenous/synthetic intronization (ESI) mutagenesis, relies on co-integration of a mutation of interest along with a selectable marker gene, the latter of which is harboured in an artificial intron adjacent to the mutation site. Cell lines generated from ESI-mutated BACs express the transgenes equivalently to the endogenous gene, and all cells efficiently splice out the synthetic intron. Thus, ESI mutagenesis provides a robust and effective single-step method with high precision and high efficiency for mutating BAC transgenes.
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Lawson, Chelsey, Patrick Batti, Prerna Singh, Madinah Suleiman Usman, Farah Bashir, Benish Alam, Gideon Asaolu, et al. "Breast Arterial Calcifications as A Predictor of Cardiovascular Risk: A Systematic Review." European Journal of Clinical Medicine 3, no. 6 (November 26, 2022): 1–5. http://dx.doi.org/10.24018/clinicmed.2022.3.6.228.
Повний текст джерелаАнотація:
Introduction: Breast arterial calcification (BAC) is considered a prediction tool for cardiovascular disease (CVD) screening as it has shown some association. The current systematic review reports the evidence on BAC detected on screening mammography and occurrence risk or likelihood of CVD among women. Methods: Three databases, including PubMed, Cochrane, and Google Scholar, were searched by three independent reviewers. Results: A total of 34,887 patients across six studies were assessed in the systematic review. The incidence of BAC on the mammogram was 10.3%. The odds or likelihood of developing CVD was higher among BAC+ women. Hypertension, hypercholesterolemia, and family history of CVD were more prevalent in BAC+ women and menopause. Conclusion: Further studies are required to develop a semi-quantitative index of BAC for CVD screening to establish BAC as a predictor of CVD further.
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Yang, Tae-Jin, Jung-Sun Kim, Ki-Byung Lim, Soo-Jin Kwon, Jin-A. Kim, Mina Jin, Jee Young Park, et al. "The KoreaBrassicaGenome Project: a Glimpse of theBrassicaGenome Based on Comparative Genome Analysis WithArabidopsis." Comparative and Functional Genomics 6, no. 3 (2005): 138–46. http://dx.doi.org/10.1002/cfg.465.
Повний текст джерелаАнотація:
A complete genome sequence provides unlimited information in the sequenced organism as well as in related taxa. According to the guidance of the Multinational Brassica Genome Project (MBGP), the Korea Brassica Genome Project (KBGP) is sequencing chromosome 1 (cytogenetically oriented chromosome #1) ofBrassica rapa. We have selected 48 seed BACs on chromosome 1 using EST genetic markers and FISH analyses. Among them, 30 BAC clones have been sequenced and 18 are on the way. Comparative genome analyses of the EST sequences and sequenced BAC clones fromBrassicachromosome 1 revealed their homeologous partner regions on theArabidopsisgenome and a syntenic comparative map betweenBrassicachromosome 1 andArabidopsischromosomes.In silicochromosome walking and clone validation have been successfully applied to extending sequence contigs based on the comparative map and BAC end sequences. In addition, we have defined the (peri)centromeric heterochromatin blocks with centromeric tandem repeats, rDNA and centromeric retrotransposons. In-depth sequence analyses of five homeologous BAC clones and anArabidopsischromosomal region reveal overall co-linearity, with 82% sequence similarity. The data indicate that theBrassicagenome has undergone triplication and subsequent gene losses after the divergence ofArabidopsisandBrassica. Based on in-depth comparative genome analyses, we propose a comparative genomics approach for conquering theBrassicagenome. In 2005 we intend to construct an integrated physical map, including sequence information from 500 BAC clones and integration of fingerprinting data and end sequence data of more than 100 000 BAC clones. The sequences have been submitted to GenBank with accession numbers: 10 204 BAC ends of the KBrH library (CW978640–CW988843); KBrH138P04, AC155338; KBrH117N09, AC155337; KBrH097M21, AC155348; KBrH093K03, AC155347; KBrH081N08, AC155346; KBrH080L24, AC155345; KBrH077A05, AC155343; KBrH020D15, AC155340; KBrH015H17, AC155339; KBrH001H24, AC155335; KBrH080A08, AC155344; KBrH004D11, AC155341; KBrH117M18, AC146875; KBrH052O08, AC155342.
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Travis, William D., Kavita Garg, Wilbur A. Franklin, Ignacio I. Wistuba, Bradley Sabloff, Masayuki Noguchi, Ryutaro Kakinuma, et al. "Evolving Concepts in the Pathology and Computed Tomography Imaging of Lung Adenocarcinoma and Bronchioloalveolar Carcinoma." Journal of Clinical Oncology 23, no. 14 (May 10, 2005): 3279–87. http://dx.doi.org/10.1200/jco.2005.15.776.
Повний текст джерелаАнотація:
Purpose To review recent advances in pathology and computed tomography (CT) of lung adenocarcinoma and bronchioloalveolar carcinoma (BAC). Methods A pathology/CT review panel of pathologists and radiologists met during a November 2004 International Association for the Study of Lung Cancer/American Society of Clinical Oncology consensus workshop in New York. The purpose was to determine if existing data was sufficient to propose modification of criteria for adenocarcinoma and BAC as newly published in the 2004 WHO Classification of Lung Tumors, and to address the pathologic/radiologic concept of diffuse/multicentric BAC. Results Solitary small, peripheral BACs have an excellent prognosis. Most lung adenocarcinomas with a BAC pattern are not pure BAC, but rather adenocarcinoma, mixed subtype with invasive patterns. This applies to tumors presenting with a diffuse/multinodular as well as solitary nodule pattern. The percent of BAC versus invasive components in lung adenocarcinomas appears to be prognostically important. However, a consensus definition of “minimally invasive” BAC with a favorable prognosis could not be achieved. While recognition of a BAC component is possible, the diagnosis of BAC with exclusion of invasive adenocarcinoma cannot be made by small biopsy or cytology specimens. Conclusion There is a need to work toward a mutual understanding and consensus between pathologists, clinicians, and researchers with the use of the term BAC versus adenocarcinoma. Future studies should make some attempt to quantitate these components and/or other features such as size of scar, size of invasive component, or pattern of invasion. Hopefully, this work will allow definition of a category of adenocarcinoma, mixed subtype with predominant BAC/minimal invasion and a favorable prognosis.
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Loberant, Norman, Vera Salamon, Nurit Carmi, and Anna Chernihovsky. "Prevalence and Degree of Breast Arterial Calcifications on Mammography: A Cross-sectional Analysis." Journal of Clinical Imaging Science 3 (September 27, 2013): 36. http://dx.doi.org/10.4103/2156-7514.119013.
Повний текст джерелаАнотація:
Objectives: The purpose of this study is to establish a database including prevalence and degree of breast arterial calcifications (BAC) in our population of women presenting for mammography. Materials and Methods: The mammograms of 1786 women over the age of 40 years were examined for the presence and degree of BAC. Statistical analysis was performed to correlate patient's age and ethnic origin with the presence and degree of BAC. Results: There was statistically significant and strong correlation between the patient's age and presence of BAC. There was also a less strong yet statistically significant correlation between patient age and degree of BAC. Regression analysis showed the likelihood of BAC at various ages. The prevalence of BAC is only 2% of women under 50 years of age; the prevalence of Grade 2-3 BAC is only 1% in women under 60 years of age. Conclusion: There is a predictable increase with age in both prevalence and degree of BAC in women. The presence of high degree BAC in women under 60 years of age or any BAC in women under 50 years of age is unusual.
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White, Robert E., Michael A. Calderwood, and Adrian Whitehouse. "Generation and precise modification of a herpesvirus saimiri bacterial artificial chromosome demonstrates that the terminal repeats are required for both virus production and episomal persistence." Journal of General Virology 84, no. 12 (December 1, 2003): 3393–403. http://dx.doi.org/10.1099/vir.0.19387-0.
Повний текст джерелаАнотація:
Herpesvirus saimiri (HVS) is the prototype gamma-2 herpesvirus, and shares considerable homology with the human gammaherpesviruses Kaposi's sarcoma-associated herpesvirus and Epstein–Barr virus. The generation of herpesvirus mutants is a key facet in the study of virus biology. The use of F-factor-based bacterial artificial chromosomes (BACs) to clone and modify the genomes of herpesviruses has enhanced the variety, precision and simplicity of mutant production. Here we describe the cloning of the genome of HVS non-transforming strain A11-S4 into a BAC. The cloning of the BAC elements disrupts open reading frame (ORF) 15 but the HVS-BAC can still replicate at levels similar to wild-type virus, and can persistently infect fibroblasts. The HVS-BAC was modified by RecA-mediated recombination initially to substitute reporter genes and also to delete the terminal repeats (TR). After deletion of the TR, the HVS-BAC fails to enter a productive virus lytic cycle, and cannot establish a persistent episomal infection when transfected into fibroblast cell lines. This shows that while ORF 15 is dispensable for virus function in vitro, the TR is required for both virus latency and lytic virus production. In addition, the HVS-BAC promises to be a valuable tool that can be used for the routine and precise production and analysis of viral mutants to further explore gammaherpesvirus biology.
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Yoon, Hee Su, Rujun Jin, Hyeon Jeong Yoon, and Kyung Chul Yoon. "Benzalkonium Chloride for Experimental Dry Eye Induction in a Rabbit Model." Journal of the Korean Ophthalmological Society 63, no. 8 (August 15, 2022): 660–68. http://dx.doi.org/10.3341/jkos.2022.63.8.660.
Повний текст джерелаАнотація:
Purpose: This study compared the clinical parameters and histological findings according to the benzalkonium chloride concentration (BAC; 0.05%, 0.1%, and 0.2%) for inducing experimental dry eye (EDE) in a rabbit model.Methods: Rabbits were divided into four groups according to the BAC concentration: untreated group, 0.05%, 0.1%, and 0.2% BAC. BAC was instilled topically in both eyes of the rabbits twice daily until they were euthanized after 14 days. Tear volume, tear break-up time (TBUT), and corneal fluorescein staining score (CFS) were measured 0, 3, 5, 7, and 14 days after treatment. After excising tissues on day 14, the conjunctival goblet cell density and corneal epithelial apoptosis were quantified.Results: The tear volume and TBUT were lower and the CFS was higher than baseline values after 14, 10, and 5 days in the 0.05, 0.1, and 0.2% BAC groups, respectively (all <i>p</i> < 0.05). At 14 days, the 0.2% BAC group showed more significant aggravation of all clinical parameters, and the 0.1% BAC group had a lower CFS (all <i>p</i> < 0.05) than the 0.05% BAC group. In all BAC groups, the conjunctival goblet cell density was lower and corneal epithelial apoptosis was significantly higher than in the untreated group (all <i>p</i> < 0.01). The conjunctival goblet cell density was lower in the 0.2% BAC group than in the 0.05% BAC group. Between-group differences in corneal epithelial apoptosis were observed in all experimental groups (all <i>p</i> < 0.01).Conclusions: Instilling BAC for 14 days induced EDE in the 0.05, 0.1, and 0.2% BAC groups. Although 0.2% BAC cannot be used for a severe EDE model, it is useful for inducing EDE in a rabbit model in a short time.
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Verster, Joris C., L. Darren Kruisselbrink, Karin A. Slot, Aikaterini Anogeianaki, Sally Adams, Chris Alford, Lizanne Arnoldy, et al. "Sensitivity to Experiencing Alcohol Hangovers: Reconsideration of the 0.11% Blood Alcohol Concentration (BAC) Threshold for Having a Hangover." Journal of Clinical Medicine 9, no. 1 (January 9, 2020): 179. http://dx.doi.org/10.3390/jcm9010179.
Повний текст джерелаАнотація:
The 2010 Alcohol Hangover Research Group consensus paper defined a cutoff blood alcohol concentration (BAC) of 0.11% as a toxicological threshold indicating that sufficient alcohol had been consumed to develop a hangover. The cutoff was based on previous research and applied mostly in studies comprising student samples. Previously, we showed that sensitivity to hangovers depends on (estimated) BAC during acute intoxication, with a greater percentage of drinkers reporting hangovers at higher BAC levels. However, a substantial number of participants also reported hangovers at comparatively lower BAC levels. This calls the suitability of the 0.11% threshold into question. Recent research has shown that subjective intoxication, i.e., the level of severity of reported drunkenness, and not BAC, is the most important determinant of hangover severity. Non-student samples often have a much lower alcohol intake compared to student samples, and overall BACs often remain below 0.11%. Despite these lower BACs, many non-student participants report having a hangover, especially when their subjective intoxication levels are high. This may be the case when alcohol consumption on the drinking occasion that results in a hangover significantly exceeds their “normal” drinking level, irrespective of whether they meet the 0.11% threshold in any of these conditions. Whereas consumers may have relative tolerance to the adverse effects at their “regular” drinking level, considerably higher alcohol intake—irrespective of the absolute amount—may consequentially result in a next-day hangover. Taken together, these findings suggest that the 0.11% threshold value as a criterion for having a hangover should be abandoned.
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Mieszkin, Sophie, Jean-Pierre Furet, G�rard Corthier, and Mich�le Gourmelon. "Estimation of Pig Fecal Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S rRNA Genetic Markers." Applied and Environmental Microbiology 75, no. 10 (March 27, 2009): 3045–54. http://dx.doi.org/10.1128/aem.02343-08.
Повний текст джерелаАнотація:
ABSTRACT The microbiological quality of coastal or river water can be affected by fecal contamination from human or animal sources. To discriminate pig fecal pollution from other pollution, a library-independent microbial source tracking method targeting Bacteroidales host-specific 16S rRNA gene markers by real-time PCR was designed. Two pig-specific Bacteroidales markers (Pig-1-Bac and Pig-2-Bac) were designed using 16S rRNA gene Bacteroidales clone libraries from pig feces and slurry. For these two pig markers, 98 to 100% sensitivity and 100% specificity were obtained when tested by TaqMan real-time PCR. A decrease in the concentrations of Pig-1-Bac and Pig-2-Bac markers was observed throughout the slurry treatment chain. The two newly designed pig-specific Bacteroidales markers, plus the human-specific (HF183) and ruminant-specific (BacR) Bacteroidales markers, were then applied to river water samples (n = 24) representing 14 different sites from the French Daoulas River catchment (Brittany, France). Pig-1-Bac and Pig-2-Bac were quantified in 25% and 62.5%, respectively, of samples collected around pig farms, with concentrations ranging from 3.6 to 4.1 log10 copies per 100 ml of water. They were detected in water samples collected downstream from pig farms but never detected near cattle farms. HF183 was quantified in 90% of water samples collected downstream near Daoulas town, with concentrations ranging between 3.6 and 4.4 log10 copies per 100 ml of water, and BacR in all water samples collected around cattle farms, with concentrations ranging between 4.6 and 6.0 log10 copies per 100 ml of water. The results of this study highlight that pig fecal contamination was not as frequent as human or bovine fecal contamination and that fecal pollution generally came from multiple origins. The two pig-specific Bacteroidales markers can be applied to environmental water samples to detect pig fecal pollution.
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Song, Junqi, Fenggao Dong, Jason W. Lilly, Robert M. Stupar, and Jiming Jiang. "Instability of bacterial artificial chromosome (BAC) clones containing tandemly repeated DNA sequences." Genome 44, no. 3 (June 1, 2001): 463–69. http://dx.doi.org/10.1139/g01-029.
Повний текст джерелаАнотація:
The cloning and propagation of large DNA fragments as bacterial artificial chromosomes (BACs) has become a valuable technique in genome research. BAC clones are highly stable in the host, Escherichia coli, a major advantage over yeast artificial chromosomes (YACs) in which recombination-induced instability is a major drawback. Here we report that BAC clones containing tandemly repeated DNA elements are not stable and can undergo drastic deletions during routine library maintenance and DNA preparation. Instability was observed in three BAC clones from sorghum, rice, and potato, each containing distinct tandem repeats. As many as 46% and 74% of the single colonies derived from a rice BAC clone containing 5S ribosomal RNA genes had insert deletions after 24 and 120 h of growth, respectively. We also demonstrated that BAC insert rearrangement can occur in the early stage of library construction and duplication. Thus, a minimum growth approach may not avoid the instability problem of such clones. The impact of BAC instability on genome research is discussed.Key words: repetitive DNA, large insert DNA library, genome research.
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Febrer, Melanie, Foo Cheung, Christopher D. Town, Steven B. Cannon, Nevin D. Young, Michael T. Abberton, Glyn Jenkins, and Dan Milbourne. "Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.)." Genome 50, no. 4 (April 2007): 412–21. http://dx.doi.org/10.1139/g07-013.
Повний текст джерелаАнотація:
White clover ( Trifolium repens L.) is a forage legume widely used in combination with grass in pastures because of its ability to fix nitrogen. We have constructed a bacterial artificial chromosome (BAC) library of an advanced breeding line of white clover. The library contains 37 248 clones with an average insert size of approximately 85 kb, representing an approximate 3-fold coverage of the white clover genome based on an estimated genome size of 960 Mb. The BAC library was pooled and screened by polymerase chain reaction (PCR) amplification using both white clover microsatellites and PCR-based markers derived from Medicago truncatula , resulting in an average of 6 hits per marker; this supports the estimated 3-fold genome coverage in this allotetraploid species. PCR-based screening of 766 clones with a multiplex set of chloroplast primers showed that only 0.5% of BAC clones contained chloroplast-derived inserts. The library was further evaluated by sequencing both ends of 724 of the clover BACs. These were analysed with respect to their sequence content and their homology to the contents of a range of plant gene, expressed sequence tag, and repeat element databases. Forty-three microsatellites were discovered in the BAC-end sequences (BESs) and investigated as potential genetic markers in white clover. The BESs were also compared with the partially sequenced genome of the model legume M. truncatula with the specific intention of identifying putative comparative-tile BACs, which represent potential regions of microsynteny between the 2 species; 14 such BACs were discovered. The results suggest that a large-scale BAC-end sequencing strategy has the potential to anchor a significant proportion of the genome of white clover onto the gene-space sequence of M. truncatula.
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Lee, Hye-Ran, Eun-Mi Eom, Yong-Pyo Lim, Jae-Wook Bang, and Dong-Hee Lee. "Construction of a garlic BAC library and chromosomal assignment of BAC clones using the FISH technique." Genome 46, no. 3 (June 1, 2003): 514–20. http://dx.doi.org/10.1139/g03-012.
Повний текст джерелаАнотація:
For molecular and cytogenetic studies, two partial bacterial artificial chromosome (BAC) libraries of the garlic cultivar Allium sativum L. 'Danyang' were constructed using high molecular weight (HMW) garlic DNA, the pBAC1SACB1 vector, and the pIndigoBAC536 vector. The average insert size of the BAC library was about 90 kb. The sequence compositions of the BAC clones were characterized by Southern hybridization with garlic genomic DNA and a repetitive sequence clone of garlic. Two BAC clones with weak signals (thus implying mostly unique sequences), GBC2-5e and GBC2-4d, were selected for FISH analysis. FISH analysis localized the GBC2-5e (~100 kb) BAC clone on the long arm of garlic chromosome 7. The other BAC clone, GBC2-4d (~110 kb), gave rise to discrete FISH signals on a mid-size early metaphase chromosome. The FISH screening with BAC clones proved to be a useful resource for molecular cytogenetic studies of garlic, and will be useful for further mapping and sequencing studies of important genes of this plant.Key words: garlic, chromosomal assignment, Allium sativum L., FISH technique, BAC library.
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Kanda, Teru, Misako Yajima, Nazmul Ahsan, Mika Tanaka, and Kenzo Takada. "Production of High-Titer Epstein-Barr Virus Recombinants Derived from Akata Cells by Using a Bacterial Artificial Chromosome System." Journal of Virology 78, no. 13 (July 1, 2004): 7004–15. http://dx.doi.org/10.1128/jvi.78.13.7004-7015.2004.
Повний текст джерелаАнотація:
ABSTRACT An Epstein-Barr virus (EBV) genome in Burkitt's lymphoma-derived cell line Akata was cloned into a bacterial artificial chromosome (BAC) vector. The BAC clone, designated AK-BAC, was rapidly and precisely modified by means of efficient homologous recombination in Escherichia coli. This system was used to produce recombinant EBVs with transgenes. An expression cassette of green fluorescent protein (GFP) was inserted into AK-BAC, and the resultant BAC clone, AK-BAC-GFP, was transfected into Akata cells. We found that transfected BAC plasmids efficiently formed episomes in EBV-positive Akata cells. Mixtures of wild-type and AK-BAC-GFP viruses were then produced and used to infect EBV-negative Akata cells. We obtained cell clones that harbored only AK-BAC-GFP but no wild-type episome. These cell clones produced infectious viruses after stimulating virus production, and the recombinant viruses of AK-BAC-GFP efficiently immortalized primary B lymphocytes. We further revised the method so that any kind of cDNA could be rapidly inserted into the unique I-PpoI site that had been artificially introduced into AK-BAC. The AK-BAC system will have a broad range of applications, such as genetic analyses of various viral gene products and development of viral vectors for human gene therapy.
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Botcherby, Marc. "Harvesting the Mouse Genome." Comparative and Functional Genomics 3, no. 4 (2002): 319–24. http://dx.doi.org/10.1002/cfg.198.
Повний текст джерелаАнотація:
The sequencing of the black 6 mouse (strain C57Bl/6) has reached an important juncture. The BAC fingerprint map is almost complete, the BACs have been endsequenced and a seven-fold coverage whole-genome shotgun has been assembled. Now the BAC-by-BAC sequencing phase is under way and in-depth comparative analysis can be carried out on regions that have been the subject of targeted sequencing. This paper reviews the progress so far and looks forward to the promises of finished sequence.
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Wu, Zhiling, and Hongbin Chen. "Comparison of invertebrate removal by traditional-BAC and pre-BAC treatment processes: verification in a full-scale drinking water treatment plant." Water Supply 18, no. 4 (September 26, 2017): 1261–69. http://dx.doi.org/10.2166/ws.2017.193.
Повний текст джерелаАнотація:
Abstract Invertebrate removal by traditional biological activated carbon (tra-BAC) and pre-BAC treatment processes was investigated in a full-scale water treatment plant. The results showed that invertebrate reproduction occurred in both BAC filters, but the invertebrate abundance in the finished water processed by tra-BAC was about 15 times greater than that processed using the pre-BAC process. In the pre-BAC process, the sand filter was placed after the BAC filter, and sand filtration removed most of the invertebrates, with an average removal efficiency of 91.1%. However, the pre-BAC filter, which was positioned behind the sedimentation tank, needed to be backwashed more frequently than the tra-BAC filter because of the high turbidity of the inlet water. The frequent backwashing reduced the biomass on the activated carbon and decreased the invertebrate reproductive rate. The results of this study are helpful for evaluating the pre-BAC treatment process in drinking water treatment plants.
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Cevik, Volkan, and Graham J. King. "Resolving the aphid resistance locus Sd-1 on a BAC contig within a sub-telomeric region of Malus linkage group 7." Genome 45, no. 5 (October 1, 2002): 939–45. http://dx.doi.org/10.1139/g02-067.
Повний текст джерелаАнотація:
Aphids cause serious physical and economic damage to most major crops throughout the world, and there is a pressing requirement to isolate genes conferring aphid resistance. The Sd-1 locus in Malus spp. (apple) confers resistance against the rosy leaf-curling aphid (Dysaphis devecta Wlk.), and was recently positioned within a 1.3-cM region on linkage group 7, flanked by molecular markers. These markers were used as a basis for development of a BAC contig spanning the locus, together with adapter-mediated amplification of flanking sequences to obtain BAC insert-end sequences, and fingerprinting of BAC clones. Approximately 800 kb of the Sd-1 genomic region was covered by 19 overlapping BACs, with an average insert size of 75150 kb. The physical genetic distance ratio was estimated at 460 kb/cM, although the distribution of recombination events was irregular with respect to estimated physical distance. Recombinant analysis and development of new markers allowed Sd-1 to be positioned within an interval of approximately 180 kb located on either of two overlapping BACs. From one of these, an insert end sequence showed a significant degree of similarity to nucleotide binding site leucine rich repeat (NBSLRR) resistance genes. Fluorescent in situ hybridization (FISH) of BAC clones within the contig enabled positioning and orientation of the locus within a euchromatic region, very close to the telomere of linkage group 7.Key words: aphid, resistance gene, apple, Malus, physical map.
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Kasuga, Ikuro, Hirotaka Nakagaki, Futoshi Kurisu, and Hiroaki Furumai. "Abundance and diversity of ammonia-oxidizing archaea and bacteria on biological activated carbon in a pilot-scale drinking water treatment plant with different treatment processes." Water Science and Technology 61, no. 12 (June 1, 2010): 3070–77. http://dx.doi.org/10.2166/wst.2010.204.
Повний текст джерелаАнотація:
The effects of different placements of rapid sand filtration on nitrification performance of BAC treatment in a pilot-scale plant were evaluated. In this plant, rapid sand filtration was placed after ozonation-BAC treatment in Process (A), while it preceded ozonation-BAC treatment in Process (B). Analysis of amoA genes of ammonia-oxidizing archaea (AOA) and bacteria (AOB) combined with nitrification potential test was conducted. BAC from Process (A) demonstrated slightly higher nitrification potential at every sampling occasion. This might be due to higher abundances of AOB on BAC from Process (A) than those on BAC from Process (B). However, AOA rather than AOB could be predominant ammonia-oxidizers in BAC treatment regardless of the position of rapid sand filtration. The highest nitrification potential was observed for BAC from both processes in February when the highest abundances of AOA-amoA and AOB-amoA genes were detected. Since rapid sand filtration was placed after BAC treatment in Process (A), residual aluminum concentration in BAC influent was higher in Process (A). However, adverse effects of aluminum on nitrification activity were not observed. These results suggest that factors other than aluminum concentration in different treatment processes could possibly have some influence on abundances of ammonia-oxidizing microorganisms on BAC.
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Shin, Se Ho, Ki Hyun Kim, Sang Seok Woo, Ju Ho Lee, Seong Hwan Kim, Jai Koo Choi, and In Suck Suh. "A Safe and Aseptic Method of Benzalkonium Chloride Dilution for Preoperative Facial Skin and Mucosa Preparation." Journal of Wound Management and Research 18, no. 2 (June 30, 2022): 114–18. http://dx.doi.org/10.22467/jwmr.2022.02012.
Повний текст джерелаАнотація:
Background: Benzalkonium chloride (BAC) is widely used as an effective antiseptic and disinfectant not only in plastic surgery, but also in everyday life. BAC has an advantage over other antiseptics used for preoperative skin preparation, such as povidone-iodine and chlorhexidine because it causes less irritation of the skin and mucosa and is less toxic. Nonetheless, when BAC is used at higher than desired concentrations, there is a possibility of adverse effects.Methods: Prior to May 2020, 10 mL of 10% BAC was mixed with 1,000 mL of distilled water and diluted to 0.1% on a weekly basis. Afterwards, the method of diluting BAC was modified; for each patient, 1.3 mL of 10% BAC and 100 mL of normal saline were mixed immediately before facial surgery.Results: From March 2007 to May 2020, 0.1% BAC was used for preoperative skin and mucosa preparation. Erroneous dilution of BAC has caused four cases of chemical burns. All the cases were attributed to human error that resulted in higher than desired concentrations of BAC in the antiseptic solution. All the affected patients suffered from first-degree and superficial second-degree chemical burns; however, they were healed uneventfully through proper wound management. Since the application of the dilution method changed since May 2020, no complications have been reported.Conclusion: We have devised a safe and aseptic method for diluting BAC. The new dilution method, which yields a constant concentration of BAC, can be used for preoperative skin and mucosa preparation without accidents.
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Matsumura, Martin E., Crystal Maksimik, Matthew W. Martinez, Michael Weiss, James Newcomb, Kenneth Harris, and Michael A. Rossi. "Breast artery calcium noted on screening mammography is predictive of high risk coronary calcium in asymptomatic women: a case control study." Vasa 42, no. 6 (November 1, 2013): 429–33. http://dx.doi.org/10.1024/0301-1526/a000312.
Повний текст джерелаАнотація:
Background: The relationship between breast artery calcification (BAC) noted on mammography and both coronary artery disease and cardiovascular risk remains controversial. Few studies have examined the clinical significance of BAC in asymptomatic women. In the present study we evaluated the relationship between BAC and coronary artery calcium (CAC) as identified by multi-slice CT scanning (MSCT). Patients and methods: Consecutive women (n = 98) with BAC noted on routine mammography but without known coronary artery disease (CAD) were assessed for CAD risk factors and had assessment of coronary calcium by MSCT. A control cohort of consecutive women who were BAC(-) (n = 104) underwent an identical assessment. Results: Women who were BAC(+) were older than those who were BAC(-); otherwise, there were no differences between the 2 groups with regard to traditional cardiac risk factors. Significantly more BAC(+) vs. BAC(-) women were found to have high risk CAC scores, defined as CAC > 400 (11.2 % vs. 1.0 %, p = 0.006). However, the rates of CAC scores of 0 were not different between the two groups (50.0 % vs. 54.8 % for BAC(+) and BAC(-) , respectively, p = 0.586). When examined in a multivariate model including the traditional risk factors of diabetes, increasing age, smoking, hyperlipidemia, and family history of CAD, the presence of BAC remained significantly associated with CAC > 400 (OR = 22.6, 95 % CI = 2.1 - 237.1). Conclusions: The presence of breast artery calcium on screening mammography was a strong independent predictor (odds ratio > 22) of high risk coronary artery calcium scores (defined as CAC > 400). The presence of BAC in those with significant CAD risk factors may warrant further evaluation.
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Wang, Dian-Hui, Huai-Ying Zhou, Chao-Hao Hu, Artem R. Oganov, Yan Zhong, and Guang-Hui Rao. "BaC: a thermodynamically stable layered superconductor." Phys. Chem. Chem. Phys. 16, no. 38 (August 18, 2014): 20780–84. http://dx.doi.org/10.1039/c4cp02781g.
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Gorantla, Padmasri, Anu Kapoor, and Jyotsna Maddury. "Mammographically Detected Breast Arterial Calcification—A Marker of Coronary Artery Disease in Women." Indian Journal of Cardiovascular Disease in Women WINCARS 02, no. 04 (December 2017): 082–85. http://dx.doi.org/10.1055/s-0038-1622963.
Повний текст джерелаАнотація:
Abstract Background and Aim Coronary artery disease (CAD) resulting from atherosclerosis is one of the major causes of mortality and morbidity worldwide and is still a leading cause of death in women older than 40 years. The gold standard for diagnosis of CAD is by coronary angiography (CAG). However, this procedure is both invasive and expensive and thus is not suitable as a screening tool in asymptomatic individuals. Like coronary calcification, atherosclerotic vascular calcification involving small-to-medium–sized arteries is also observed in the breast. Modern mammographic equipment is very sensitive in the detection of microcalcifications. The aim of this study is to study the relationship between mammographically detected breast arterial calcification (BAC) and CAD and to evaluate the role of BAC as a marker for CAD. Methods Twenty female patients older than 40 years who had undergone CAG for suspected CAD were included in the study. Screening mammograms were performed and analyzed for the presence of BAC. The results were analyzed for correlation between severity of BAC and CAD. Results Twenty patients with mean age of 56.45 (age range: 40–68) were evaluated. BACs were found in 60% of these cases with a peak age group of 56 to 60 years. BAC was bilateral (92%) in most cases. CAG reports were positive for CAD in 45% of patients. The sensitivity of BAC in predicting CAD was 77.7% with a specificity of 54.5%, positive predictive value (PPV) of 58.3%, and negative predictive value (NPV) of 75%. Conclusion Screening mammography has a potential to serve as a noninvasive tool for early detection of CAD in asymptomatic women. Larger population-based studies with controls will be required to establish the utility of this screening tool.
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Kim, Jeong-Soon, Kevin L. Childs, M. Nurul Islam-Faridi, Monica A. Menz, Robert R. Klein, Patricia E. Klein, H. James Price, John E. Mullet, and David M. Stelly. "Integrated karyotyping of sorghum by in situ hybridization of landed BACs." Genome 45, no. 2 (April 1, 2002): 402–12. http://dx.doi.org/10.1139/g01-141.
Повний текст джерелаАнотація:
The reliability of genome analysis and proficiency of genetic manipulation are increased by assignment of linkage groups to specific chromosomes, placement of centromeres, and orientation with respect to telomeres. We have endeavored to establish means to enable these steps in sorghum (Sorghum bicolor (L.) Moench), the genome of which contains ca. 780 Mbp spread across n = 10 chromosomes. Our approach relies on fluorescence in situ hybridization (FISH) and integrated structural genomic resources, including large-insert genomic clones in bacterial artificial chromosome (BAC) libraries. To develop robust FISH probes, we selected sorghum BACs by association with molecular markers that map near the ends of linkage groups, in regions inferred to be high in recombination. Overall, we selected 22 BACs that encompass the 10 linkage groups. As a prelude to development of a multiprobe FISH cocktail, we evaluated BAC-derived probes individually and in small groups. Biotin- and digoxygenin-labeled probes were made directly from the BAC clones and hybridized in situ to chromosomes without using suppressive unlabelled C0t-1 DNA. Based on FISH-signal strength and the relative degree of background signal, we judged 19 BAC-derived probes to be satisfactory. Based on their relative position, and collective association with all 10 linkage groups, we chose 17 of the 19 BACs to develop a 17-locus probe cocktail for dual-color detection. FISH of the cocktail allowed simultaneous identification of all 10 chromosomes. The results indicate that linkage and physical maps of sorghum allow facile selection of BAC clones according to position and FISH-signal quality. This capability will enable development of a high-quality molecular cytogenetic map and an integrated genomics system for sorghum, without need of chromosome flow sorting or microdissection. Moreover, transgeneric FISH experiments suggest that the sorghum system might be applicable to other Gramineae.Key words: integrated karyotyping, FISH, sorghum, BAC.
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Yang, Zhong-Nan, and T. Erik Mirkov. "Isolation of large terminal sequences of BAC inserts based on double-restriction-enzyme digestion followed by anchored PCR." Genome 43, no. 2 (March 15, 2000): 412–15. http://dx.doi.org/10.1139/g99-120.
Повний текст джерелаАнотація:
Isolation of the terminal portions of genomic DNA cloned in bacterial artificial chromosomes (BACs) is an important step in map-based cloning, and several methods have been developed. Here, we present a new method based on double-restriction-enzyme digestion followed by anchored PCR. BAC DNA was digested with two enzymes: NotI and one of four enzymes (EcoRV, HpaI, StuI, or XmnI) that produce blunt termini. After dephosphorylation, these digestions were ligated to NotI- and EcoRV-digested pMSK, a new cloning vector developed in this work that is derived from pBluescript SK(+). PCR products representing the left- and right-terminal sequences of BAC inserts were obtained using a primer complementary to pMSK and a primer complementary to sequences in either the left arm or the right arm of the BAC vector pBeloBAC11. We have tested this method with 15 different BAC clones, and PCR products representing both the left- and right-terminal sequences have been obtained from all 15 BAC clones. This method is simple, fast, reproducible, and uses the same set of primers for any restriction enzyme used. With some modifications, it can also be used for isolating the terminal portions of genomic DNA cloned in yeast artificial chromosomes and P1-derived artificial chromosomes. Key words: BAC, anchored PCR, terminal sequence isolation, chromosome walk.
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Adhikari, Mandira Pradhananga, and Sandeep Sharma Lamsal. "Silver(I) Ion Removal Efficiency of Activated Carbon Prepared from Terminalia-bellerica (Barro) Seed Stone." Journal of Nepal Chemical Society 42, no. 1 (March 1, 2021): 89–98. http://dx.doi.org/10.3126/jncs.v42i1.35340.
Повний текст джерелаАнотація:
Phosphoric acid-activated Terminalia-bellerica (Barro) seed stone powder was carbonized in a muffle furnace at three different temperatures (300, 400, and 500oC). The activated carbons (BAC-300, BAC-400, and BAC-500) were characterized by using Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), methylene blue number, and iodine number. The iodine number (357 mg/g) and specific surface area (537 m2/g) were a maximum for BAC-400. The BACs followed Langmuir adsorption isotherm and the maximum methylene blue adsorption capacity was 212.77 mg/g. The silver ion removal efficiency was a maximum at pH 6, 3 mg/L of adsorbent dose, and 20 mg/L of silver ion concentration. The BAC-400 could adsorb 40 % of silver ion within 5 mins with the initial Ag(I) ion concentration of 20 mg/L and an adsorbent dose of 1 mg/L. The percentage of adsorption enhanced to 100% with the increment of adsorbent doseto3 g/L.The adsorption kinetics of silver (I) ion on BAC-400 was well fitted to pseudo-second-order kinetics suggesting the chemisorption of silver ions. All the results attributed that low-cost viable adsorbent can be prepared from Barro seed stone for the efficient removal of silver ion from aqueous solution.
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Tischer, B. Karsten, and Benedikt B. Kaufer. "Viral Bacterial Artificial Chromosomes: Generation, Mutagenesis, and Removal of Mini-F Sequences." Journal of Biomedicine and Biotechnology 2012 (2012): 1–14. http://dx.doi.org/10.1155/2012/472537.
Повний текст джерелаАнотація:
Maintenance and manipulation of large DNA and RNA virus genomes had presented an obstacle for virological research. BAC vectors provided a solution to both problems as they can harbor large DNA sequences and can efficiently be modified using well-established mutagenesis techniques inEscherichia coli. Numerous DNA virus genomes of herpesvirus and pox virus were cloned into mini-F vectors. In addition, several reverse genetic systems for RNA viruses such as members ofCoronaviridaeandFlaviviridaecould be established based on BAC constructs. Transfection into susceptible eukaryotic cells of virus DNA cloned as a BAC allows reconstitution of recombinant viruses. In this paper, we provide an overview on the strategies that can be used for the generation of virus BAC vectors and also on systems that are currently available for various virus species. Furthermore, we address common mutagenesis techniques that allow modification of BACs from single-nucleotide substitutions to deletion of viral genes or insertion of foreign sequences. Finally, we review the reconstitution of viruses from BAC vectors and the removal of the bacterial sequences from the virus genome during this process.
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Kunec, Dusan, Larry A. Hanson, Sandra van Haren, I. F. Nieuwenhuizen, and Shane C. Burgess. "An Overlapping Bacterial Artificial Chromosome System That Generates Vectorless Progeny for Channel Catfish Herpesvirus." Journal of Virology 82, no. 8 (January 30, 2008): 3872–81. http://dx.doi.org/10.1128/jvi.02152-07.
Повний текст джерелаАнотація:
ABSTRACT Herpesviruses are important pathogens of humans and other animals. Herpesvirus infectious clones that can reconstitute phenotypically wild-type (wt) virus are extremely valuable tools for elucidating the roles of specific genes in virus pathophysiology as well as for making vaccines. Ictalurid herpesvirus 1 (channel catfish herpesvirus [CCV]) is economically very important and is the best characterized of the herpesviruses that occur primarily in bony fish and amphibians. Here, we describe the cloning of the hitherto recalcitrant CCV genome as three overlapping subgenomic bacterial artificial chromosomes (BACs). These clones allowed us to regenerate vectorless wt CCVs with a phenotype that is indistinguishable from that of the wt CCV from which the BACs were derived. To test the recombinogenic systems, we next used the overlapping BACs to construct a full-length CCV BAC by replacing the CCV ORF5 with the BAC cassette and cotransfecting CCO cells. The viral progeny that we used to transform Escherichia coli and the resulting BAC had only one of the 18-kb terminal repeated regions. Both systems suggest that one of the terminal repeat regions is lost during the replicative stage of the CCV life cycle. We also demonstrated the feasibility of introducing a targeted mutation into the CCV BAC infectious clone by constructing a CCV ORF12 deletion mutant and showed that ORF12 encodes a nonessential protein for virus replication. This is the first report of the generation of an infectious BAC clone of a member of the fish and amphibian herpesviruses and its use to generate recombinants.
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Nishijima, Wataru, and Mitsumasa Okada. "Particle separation as a pretreatment of an advanced drinking water treatment process by ozonation and biological activated carbon." Water Science and Technology 37, no. 10 (May 1, 1998): 117–24. http://dx.doi.org/10.2166/wst.1998.0390.
Повний текст джерелаАнотація:
The role of particle separation on the performance of ozone-biological activated carbon (BAC) was evaluated based on the analyses of the fate of organic substances in the process. Pilot plant studies were carried out using eutrophic lake water as raw water. The ozonation not only converted refractory organic matter into biodegradable matter but also particulate organic carbon (POC) into dissolved organic carbon (DOC). Total decrease in adsorbable and non biodegradable DOC fraction (ADOC) after ozonation was only 16% of the influent into the biofiltration process followed by ozonation. However, the ozone-BAC process before membrane separation could reduce organic loading to membrane system. The smaller loading to microfiltration will result in long intervals of back washing and less frequent membrane fouling. Membrane separation before ozonation removed not only POC but also a part of DOC and could prevent dissolution of POC during ozonation. The decreases in ADOC by membrane and ozonation were 20% and 37% of the influent ADOC, respectively. The total decrease in ADOC for membrane process followed by ozonation was 57%. The separation of particulate matter will decrease loading of ADOC onto BAC significantly and, therefore, will extend service life of BAC.