Добірка наукової літератури з теми "Isopropyl β-D-1 thiogalactopyranoside"

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Статті в журналах з теми "Isopropyl β-D-1 thiogalactopyranoside"

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Sun, Huigang, Xiaomei Bie, Fengxia Lu, Yaping Lu, Yundailai Wu, and Zhaoxin Lu. "Enhancement of surfactin production of Bacillus subtilis fmbR by replacement of the native promoter with the Pspac promoter." Canadian Journal of Microbiology 55, no. 8 (August 2009): 1003–6. http://dx.doi.org/10.1139/w09-044.

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Анотація:
Bacillus subtilis fmbR-1 was obtained from wild-type B. subtilis fmbR by replacement of the native promoter of the surfactin operon with the inducible promoter Pspac. The recombinant B. subtilis fmbR-1 produced more antibacterial substances than the wild-type strain. The overproducing phenotype was related to the enhancement of antagonistic activities against Bacillus cereus . HPLC peaks of surfactin for recombinant fmbR-1 showed patterns of lipopeptides similar to those of the wild-type strain, and surfactin production of the recombinant fmbR-1 was up to about fivefold greater than that of the wild-type strain without induction by isopropyl β-d-1-thiogalactopyranoside. However, the production of surfactin increased to about 10-fold more than that of the wild-type strain when it was induced by isopropyl β-d-1-thiogalactopyranoside.
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Lee, Sung Kuk, Howard H. Chou, Brian F. Pfleger, Jack D. Newman, Yasuo Yoshikuni, and Jay D. Keasling. "Directed Evolution of AraC for Improved Compatibility of Arabinose- and Lactose-Inducible Promoters." Applied and Environmental Microbiology 73, no. 18 (July 20, 2007): 5711–15. http://dx.doi.org/10.1128/aem.00791-07.

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ABSTRACT Synthetic biological systems often require multiple, independently inducible promoters in order to control the expression levels of several genes; however, cross talk between the promoters limits this ability. Here, we demonstrate the directed evolution of AraC to construct an arabinose-inducible (PBAD) system that is more compatible with IPTG (isopropyl-β-d-1-thiogalactopyranoside) induction of a lactose-inducible (Plac) system. The constructed system is 10 times more sensitive to arabinose and tolerates IPTG significantly better than the wild type. Detailed studies indicate that the AraC dimerization domain and C terminus are important for the increased sensitivity of AraC to arabinose.
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HUBCHYK, K. A., R. N. BIRUKOU, А. М. НLUSHEN, I. S. KAZLOUSKI, and A. A. KASTSIANEVICH. "GENETIC ENGINEERING OF STRAIN ESCHERICHIA COLI BL21.BT1 CAPABLE TO SYNTHESIZE RECOMBINANT BETA-1,3-NACETYLGLUCOSAMINE TRANSFERASE." Микробные биотехнологии: фундаментальные и прикладные аспекты 13 (October 21, 2021): 52–65. http://dx.doi.org/10.47612/2226-3136-2021-13-52-65.

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A strain of Escherichia coli BL21.Bt1, a producer of the recombinant beta-1,3-N-acetylglucosamine transferase Bacillus thuringiensis BIM B-180, has been constructed. The cultivation conditions of the producer strain are optimized: the initial pH value of the nutrient medium is 7.2; cultivation temperature after induction – 20 °C; constant stirring at an intensity of 200 rpm; the use of 1 mM isopropyl-β-D-1-thiogalactopyranoside as an inducer; introduction of 10 mM lactose 3 h after induction. It was shown that the yield of the target enzymatic protein after 24 h of cultivation of the E. coli BL21.Bt1 strain under optimized conditions reaches 63 μg/ml.
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Sitasiwi, Agung Janika, Wayan Tunas Artama, Agung Budiyanto, and Edy Dharmana. "MOLECULAR EXPRESSION OF WINGLESS-TYPE MMTV INTEGRATION SITE FAMILY MEMBER 4 GENE USING Escherichia coli BL21." Jurnal Kedokteran Hewan - Indonesian Journal of Veterinary Sciences 11, no. 1 (April 7, 2017): 11–14. http://dx.doi.org/10.21157/j.ked.hewan.v11i1.5891.

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Анотація:
This research was conducted to find out the Wnt4 recombinant proteins which expressed by Escherichia coli (E. coli) BL21 carrying the recombinant DNA wnt4 (E. coli transformation). Research materials were E. coli BL21 transformation and E. coli BL21 non-transformation (negative control). The expression of recombinant protein was conducted by culturing E. coli for 24 hours in Luria-Bertani (LB) media with isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. Recombinant protein was isolated by sonication of pellet bacteria. Protein analysis performed by 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed that recombinant protein with a molecular weight of 33 kDa has been expressed by E. coli BL21 transformation successfully.
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Muttar, Arafat, and Intesar Tarik Numan. "Cloning & expression of SAK enzyme from Staphylococcus aureus in E. coli BL21-CodonPlus." Journal of Medicine and Life 15, no. 6 (June 2022): 768–71. http://dx.doi.org/10.25122/jml-2021-0335.

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Staphylokinase (SAK), also known as staphylococcal fibrinolysin, is a protein with a molecular mass of about 15 kDa produced by Staphylococcus aureus. Staphylokinase is synthesized in the late exponential phase, similar to streptokinase. The current study identified and predicted the protein SAK from Staphylococcus aureus. SAK is a fibrinolytic enzyme of the third generation that acts as an indirect activator of plasminogen. The current study cloned and expressed SAK protein isolated from Staphylococcus aureus and used in the form of a grid for enhancement of SAK Catalyst with PCR, disengagement, and change into articulation vector PET24b(+). The recombinant plasmid was changed into E. coli strain BL21 (codon additionally to 440) acceptance with isopropyl β-D-1-thiogalactopyranoside (IPTG).
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Shafat, Zoya, Abu Hamza, Farah Deeba, Md Imam Faizan, Nazim Khan, Asimul Islam, Anwar Ahmed, Salman F. Alamery, and Shama Parveen. "Optimization of parameters for expression and purification of G glycoprotein ectodomain of respiratory syncytial virus." Future Virology 15, no. 4 (April 2020): 225–35. http://dx.doi.org/10.2217/fvl-2019-0157.

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Анотація:
Aim: G glycoprotein ectodomain (Ge) of BA genotype of group B respiratory syncytial virus was expressed and purified to achieve maximum yield of the protein. Materials & methods: We optimized different parameters like strains, temperature, inducer concentration and post induction time period for efficient protein expression in Escherichia coli. The protein was purified using affinity chromatography and confirmed by western blotting. Results: It was concluded that a 5-h induction with 0.75 mM isopropyl β-D-1-thiogalactopyranoside at 37°C in BL21(DE3) cells was the most favorable condition for maximal protein expression. The far-UV circular dichroism spectroscopy suggested that it is an α-helical protein. Conclusion: The purified Ge protein can be characterized by antigenic and biophysical methods in future studies, which will probably assist in vaccine development.
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Grinanda, Dita, and Takashi Hirasawa. "Effectiveness of the Bacillus subtilis genome-reduced strain as an ethanol production host." Bioscience, Biotechnology, and Biochemistry 86, no. 4 (January 31, 2022): 543–51. http://dx.doi.org/10.1093/bbb/zbac017.

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ABSTRACT We investigated the performance of a genome-reduced strain of Bacillus subtilis MGB874, whose 0.87 Mbp of genomic DNA was cumulatively deleted, as an ethanol production host. A recombinant strain A267_EtOH was constructed by introducing the pdc and adhB genes from Zymomonas mobilis, both of which were expressed from an isopropyl-β-d-1-thiogalactopyranoside-inducible spac promoter, into the A267 strain, a tryptophan prototrophic derivative of the MGB874 with disruption of metabolic pathways for producing lactic acid, acetic acid, and acetoin. Focusing on the stationary phase in fed-batch fermentation, 1.6 g L−1 ethanol was produced by the A267_EtOH strain after 144 h. Moreover, its ethanol production further increased by approximately 3.7-fold (5.9 g L−1) at 80 h through replacing the spac promoter for expressing pdc and adhB genes with the lytR promoter and the yield was about 112%. These results indicate that the MGB874 is an effective host for ethanol production during the stationary phase.
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Zhen-Hui, Song, Guo Wan-Zhu, and Zhang Ying-Jun. "Prokaryotic expression of nucleoprotein gene of Transmissible gastroenteritis virus and development of ELISA based on the expressed protein." Chinese Journal of Agricultural Biotechnology 6, no. 3 (December 2009): 249–52. http://dx.doi.org/10.1017/s1479236209990520.

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AbstractThe recombinant PET-N plasmid, which includes the N gene of the Transmissible gastroenteritis virus (TGEV), was transformed into Escherichia coli BL21 (DE3), and induced at 37°C with 1.0 mmol/l IPTG (isopropyl β-d-1-thiogalactopyranoside). An indirect enzyme-linked immunosorbent assay (ELISA) for detecting the TGEV nucleocapsid protein antibody was developed after the reactogenicity of the recombinant protein was demonstrated by Western blot. The operating conditions for the ELISA, an antigen concentration of 15 μg/ml, serum dilution of 1:40, blocking solution of 0.5% fetal bovine serum (FBS), serum sample incubation for 90 min, a concentration of horseradish peroxidase (HRP)-spa of 1:5000, incubated for 60 min, incubation of the substrate at room temperature for 10 min, and a cutoff value for the ELISA OD450⩾0.35, were carried out using a checkerboard titration method. The sensitivity and specificity of this method relative to the Svanova TGEV/PRCV antibody diagnosis kit were 93.5% and 93.8%, respectively.
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Lipničanová, Sabina, Barbora Legerská, Daniela Chmelová, Miroslav Ondrejovič, and Stanislav Miertuš. "Optimization of an Inclusion Body-Based Production of the Influenza Virus Neuraminidase in Escherichia coli." Biomolecules 12, no. 2 (February 19, 2022): 331. http://dx.doi.org/10.3390/biom12020331.

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Анотація:
Neuraminidase (NA), as an important protein of influenza virus, represents a promising target for the development of new antiviral agents for the treatment and prevention of influenza A and B. Bacterial host strain Escherichia coli BL21 (DE3)pLysS containing the NA gene of the H1N1 influenza virus produced this overexpressed enzyme in the insoluble fraction of cells in the form of inclusion bodies. The aim of this work was to investigate the effect of independent variables (propagation time, isopropyl β-d-1-thiogalactopyranoside (IPTG) concentration and expression time) on NA accumulation in inclusion bodies and to optimize these conditions by response surface methodology (RSM). The maximum yield of NA (112.97 ± 2.82 U/g) was achieved under optimal conditions, namely, a propagation time of 7.72 h, IPTG concentration of 1.82 mM and gene expression time of 7.35 h. This study demonstrated that bacterially expressed NA was enzymatically active.
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Jie, Jing, Xiao Chu, Dan Li, and Zhaoqing Luo. "A set of shuttle plasmids for gene expression in Acinetobacter baumannii." PLOS ONE 16, no. 2 (February 10, 2021): e0246918. http://dx.doi.org/10.1371/journal.pone.0246918.

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Анотація:
Infections caused by the emerging opportunistic bacterial pathogen Acinetobacter baumannii are occurring at increasingly alarming rates, and such increase in incidence is further compounded by the development of wide spread multidrug-resistant strains. Yet, our understanding of its pathogenesis and biology remains limited which can be attributed in part to the scarce of tools for molecular genetic analysis of this bacterium. Plasmids based on pWH1277 originally isolated from Acinetobacter calcoaceticus are the only vehicles currently available for ectopic gene expression in Acinetobacter species, which restricts experiments that require simultaneous analysis of multiple genes. Here, we found that plasmids of the IncQ group are able to replicate in A. baumannii and can stably co-reside with derivatives of pWH1277. Furthermore, we have constructed a series of four plasmids that allow inducible expression of Flag-tagged proteins in A. baumannii by arabinose or isopropyl β-d-1-thiogalactopyranoside. Together with constructs previously developed, these plasmids will accommodate the need in genetic analysis of this increasingly important pathogen.
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Тези доповідей конференцій з теми "Isopropyl β-D-1 thiogalactopyranoside"

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Wu, Yi-Hsuan, Wen-Tai Chiu, Meng-Jia Lian та Chih-Ling Huang. "Scanned Laser Pico-Projection Image Processing for Cancer-Like Cells Induced by Isopropyl β-D-1-thiogalactopyranoside". У the 2018 8th International Conference. New York, New York, USA: ACM Press, 2018. http://dx.doi.org/10.1145/3208955.3208973.

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