Academic literature on the topic 'Human brain cDNA library'

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Journal articles on the topic "Human brain cDNA library"

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Adams, Mark D., M. Bento Soares, Anthony R. Kerlavage, Chris Fields, and J. Craig Venter. "Rapid cDNA sequencing (expressed sequence tags) from a directionally cloned human infant brain cDNA library." Nature Genetics 4, no. 4 (August 1993): 373–80. http://dx.doi.org/10.1038/ng0893-373.

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Xu, Lei, Jingqi Li, Li Liu, Lixia Lu, Jingxia Gao, and Xueli Li. "Construction of equalized short hairpin RNA library from human brain cDNA." Journal of Biotechnology 128, no. 3 (February 20, 2007): 477–85. http://dx.doi.org/10.1016/j.jbiotec.2006.11.013.

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Nobis, William, Xiaoning Ren, Steven P. Suchyta, Thomas R. Suchyta, Adroaldo J. Zanella, and Paul M. Coussens. "Development of a porcine brain cDNA library, EST database, and microarray resource." Physiological Genomics 16, no. 1 (December 16, 2003): 153–59. http://dx.doi.org/10.1152/physiolgenomics.00099.2003.

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Recent developments in expressed sequence tag (EST) and cDNA microarray technology have had a dramatic impact on the ability of scientists to study responses of thousands of genes to internal and external stimuli. In neurobiology, studies of the human brain have been expanding rapidly by use of functional genomics techniques. To enhance these studies and allow use of a porcine brain model, a normalized porcine brain cDNA library (PBL) has been generated and used as a base for EST discovery and microarray generation. In this report, we discuss initial sequence analysis of 965 clones from this resource. Our data revealed that library normalization successfully reduced the number of clones representing highly abundant cDNA species and overall clone redundancy. Cluster analysis revealed over 800 unique cDNA species representing a redundancy rate for the normalized library of 6.9% compared with 29.4% before normalization. Sequence information, BLAST results, and TIGR cluster matches for these ESTs are publicly available via a web-accessible database ( http://nbfgc.msu.edu ). A cDNA microarray was created using 877 unique porcine brain EST amplicons spotted in triplicate on glass slides. This microarray was assessed by performing a series of experiments designed to test hybridization efficiency and false-positive rate. Our results indicate that the PBL cDNA microarray is a robust tool for studies of brain gene expression using swine as a model system.
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Gong, Yuewen, Manna Zhang, Li Cui, and Gerald Y. Minuk. "Sequence and chromosomal assignment of a human novel cDNA: similarity to gamma-aminobutyric acid transporter." Canadian Journal of Physiology and Pharmacology 79, no. 12 (December 1, 2001): 977–84. http://dx.doi.org/10.1139/y01-082.

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Gamma-aminobutyric acid (GABA) is the predominant inhibitory neurotransmitter in the mammalian brain. Although initially thought to be confined to the central nervous system, GABAergic activity has also been described in other tissues throughout the body. In the present study, we report the cloning and localization of human GABA transporter cDNA and document its expression in various human tissues. A human liver cDNA library was initially screened by a 32P-labeled murine brain GABA transporter 3 (GAT-3) cDNA probe, and full-length cDNA was cloned by employing Marathon-Ready(tm) human kidney cDNA. The human GABA transporter cDNA encoded a 569 amino acid hydrophobic protein with 12 transmembrane domains (TMs). Search of published sequences revealed high homology with rat GAT-2, murine GAT-3 cDNA, human solute carrier family 6 member 13 (SLC6A13), and a human peripheral betaine/GABA transporter. Northern blot analyses demonstrated that the human GABA transporter is expressed strongly in the kidney and to a lesser extent in the liver and brain. The sequence was well matched with human chromosome 12p13.3, suggesting the human GABA transporter contains 14 exons. The above findings confirm the existence of and further characterize a specific GABA transporter in human tissues.Key words: sequence, chromosome, GABA, GABA transporter.
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MICHEL, UWE, BORIS KALLMANN, PETER RIECKMANN, and DIRK ISBRANDT. "UM 9(5)h and UM 9(5)p, human and porcine noncoding transcripts with preferential expression in the cerebellum." RNA 8, no. 12 (December 2002): 1538–47. http://dx.doi.org/10.1017/s1355838202028042.

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We compared the gene expression patterns of fetal and adult porcine brains and identified a sequence tag that was more abundant in adult than in fetal brain. The RNA corresponding to the sequence tag has the highest expression level in adult cerebellum. Lower expression levels of the transcript were found in adult cerebrum, pituitary, and uterus, as well as in fetal brain, heart, intestine, kidney, and liver. The sequence tag was used to screen a cDNA library from adult porcine brain. Two independent clones of 2,273 nt and 1,701 nt were isolated. The shorter cDNA is a 5′-truncated form of the longer clone, and both clones have almost identical sequences with multiple start and stop codons in all three reading frames. Screening of two different human brain cDNA libraries with porcine cDNA probes resulted in four overlapping cDNA fragments, which were assembled to one contig of 2,336 nt in length. Like noncoding RNAs, the porcine and human sequences have no common conserved open reading frame and share stretches of high homology interrupted by stretches with almost no homology. The human and porcine RNAs were named UM 9(5)h and UM 9(5)p, respectively. They are part of larger transcripts, which are transcribed from single-copy genes, they have very similar tissue distributions, and their sequences are colinear with the respective genomic fragment.
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KIKONYOGO, Alexandra, and Regina PIETRUSZKO. "Aldehyde dehydrogenase from adult human brain that dehydrogenates γ-aminobutyraldehyde: purification, characterization, cloning and distribution." Biochemical Journal 316, no. 1 (May 15, 1996): 317–24. http://dx.doi.org/10.1042/bj3160317.

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Enzyme purification and characterization, cDNA cloning and Northern blot analysis were the techniques utilized during this investigation to determine the identity and occurrence of the aldehyde dehydrogenase that metabolizes γ-aminobutyraldehyde in adult human brain. The purification yielded one major protein which was active with γ-aminobutyraldehyde. It had the physicochemical and kinetic properties of the human liver E3 isoenzyme of aldehyde dehydrogenase (EC 1.2.1.3), and also interacted with an anti-(liver E3 isoenzyme) antibody. Tryptic peptides derived from the purified brain protein matched the amino acid sequence of the liver E3 isoenzyme. Employing liver E3 cDNA, a human cerebellar cDNA library was screened and a 2.0 kb cDNA fragment was isolated. The cerebellar cDNA yielded a derived primary structure which differed from the liver E3 amino acid sequence by a single serine-to-cysteine substitution at position 88 (position 84 in the liver sequence). Thus the γ-aminobutyraldehyde-metabolizing enzyme from human brain can be identified as E3′, a variant of the E3 isoenzyme. The catalytic properties of the brain variant were indistinguishable from those of E3, and so the functional importance of this variant is at present unknown. The distribution of this enzyme in brain was investigated by Northern blot analysis, which demonstrated the presence of E3′ mRNA in all regions of the human brain. mRNA levels were variable in the different brain areas, with the highest levels in the spinal cord and the lowest in the occipital pole.
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Thomson, S. A. M., E. Kennerly, N. Olby, J. R. Mickelson, D. E. Hoffmann, P. J. Dickinson, G. Gibson, and M. Breen. "Microarray Analysis of Differentially Expressed Genes of Primary Tumors in the Canine Central Nervous System." Veterinary Pathology 42, no. 5 (September 2005): 550–58. http://dx.doi.org/10.1354/vp.42-5-550.

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The pathophysiologic similarities of many human and canine cancers support the role of the domestic dog as a model for brain tumor research. Here we report the construction of a custom canine brain-specific cDNA microarray and the analysis of gene expression patterns of several different types of canine brain tumor The microarray contained 4000 clones from a canine brain specific cDNA library including 2161 clones that matched known genes or expressed sequence tags (ESTs) and 25 cancer-related genes. Our study included 16 brain tumors (seven meningiomas, five glial tumors, two ependymomas, and two choroid plexus papillomas) from a variety of different dog breeds. We identified several genes previously found to be differentially expressed in human brain tumors. This suggests that human and canine brain tumors share a common pathogenesis. In addition, we also found differentially expressed genes unique to either meningiomas or the glial tumors. This report represents the first global gene expression analysis of different types of canine brain tumors by cDNA microarrays and might aid in the identification of potential candidate genes involved in tumor formation and progression.
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Middlemas, D. S., R. A. Lindberg, and T. Hunter. "trkB, a neural receptor protein-tyrosine kinase: evidence for a full-length and two truncated receptors." Molecular and Cellular Biology 11, no. 1 (January 1991): 143–53. http://dx.doi.org/10.1128/mcb.11.1.143-153.1991.

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We have screened an adult rat cerebellar cDNA library in search of novel protein tyrosine-kinase (PTK) cDNAs. A cDNA for a putative PTK, trkB, was cloned, and its sequence indicates that it is likely to be derived from a gene for a ligand-regulated receptor closely related to the human trk oncogene. Northern (RNA) analysis showed that the trkB gene is expressed predominantly in the brain and that trkB expresses multiple mRNAs, ranging from 0.7 to 9 kb. Hybridization of cerebral mRNAs with a variety of probes indicates that there are mRNAs encoding truncated trkB receptors. Two additional types of cDNA were isolated, and their sequences are predicted to encode two distinct C-terminally truncated receptors which have the complete extracellular region and transmembrane domain, but which differ in their short cytoplasmic tails.
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Middlemas, D. S., R. A. Lindberg, and T. Hunter. "trkB, a neural receptor protein-tyrosine kinase: evidence for a full-length and two truncated receptors." Molecular and Cellular Biology 11, no. 1 (January 1991): 143–53. http://dx.doi.org/10.1128/mcb.11.1.143.

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We have screened an adult rat cerebellar cDNA library in search of novel protein tyrosine-kinase (PTK) cDNAs. A cDNA for a putative PTK, trkB, was cloned, and its sequence indicates that it is likely to be derived from a gene for a ligand-regulated receptor closely related to the human trk oncogene. Northern (RNA) analysis showed that the trkB gene is expressed predominantly in the brain and that trkB expresses multiple mRNAs, ranging from 0.7 to 9 kb. Hybridization of cerebral mRNAs with a variety of probes indicates that there are mRNAs encoding truncated trkB receptors. Two additional types of cDNA were isolated, and their sequences are predicted to encode two distinct C-terminally truncated receptors which have the complete extracellular region and transmembrane domain, but which differ in their short cytoplasmic tails.
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Gérard, C. M., C. Mollereau, G. Vassart, and M. Parmentier. "Molecular cloning of a human cannabinoid receptor which is also expressed in testis." Biochemical Journal 279, no. 1 (October 1, 1991): 129–34. http://dx.doi.org/10.1042/bj2790129.

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A cDNA clone encoding a receptor protein which presents all the characteristics of a guanine-nucleotide-binding protein (G-protein)-coupled receptor was isolated from a human brain stem cDNA library. The probe used (HGMP08) was a 600 bp DNA fragment amplified by a low-stringency PCR, using human genomic DNA as template and degenerate oligonucleotide primers corresponding to conserved sequences amongst the known G-protein-coupled receptors. The deduced amino acid sequence encodes a protein of 472 residues which shares 97.3% identity with the rat cannabinoid receptor cloned recently [Matsuda, Lolait, Brownstein, Young & Bronner (1990) Nature (London) 346, 561-564]. Abundant transcripts were detected in the brain, as expected, but lower amounts were also found in the testis. The same probe was used to screen a human testis cDNA library. The cDNA clones obtained were partially sequenced, demonstrating the identity of the cannabinoid receptors expressed in both tissues. Specific binding of the synthetic cannabinoid ligand [3H]CP55940 was observed on membranes from Cos-7 cells transfected with the recombinant receptor clone. In stably transfected CHO-K1 cell lines, cannabinoid agonists mediated a dose-dependent and stereoselective inhibition of forskolin-induced cyclic AMP accumulation. The ability to express the human cannabinoid receptor in mammalian cells should help in developing more selective drugs, and should facilitate the search for the endogenous cannabinoid ligand(s).
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Dissertations / Theses on the topic "Human brain cDNA library"

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Marson, Lorena. "Phage-display epitope library development for biomarkers identification in autoimmune diseases of the Central Nervous System." Doctoral thesis, Università degli studi di Trieste, 2012. http://hdl.handle.net/10077/7405.

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2010/2011
The principal aim of my PhD was the setting of a protocol for the creation of phage libraries to display cDNA fragments encoding real ORF sequences, that could correspond to potential epitopes. A similar phage display library contains all the potential ORF repertoire of a cell or tissue. This tool can be specially used in the study of autoimmune diseases to perform different kind of analysis, such as the identification of epitopes involved in pathological reaction, the comparison between healthy and pathological conditions, or between different pathological conditions. A complex protocol was developed. It provides for: cDNA normalization, cDNA fragmentation to obtain peptides with useful size, and ORF enrichment to obtain really coding fragments. With this system we have created a epitopes library from Human brain mRNA.
Il principale obiettivo del mio lavoro di ricerca è la messa a punto di un protocollo per la costruzione di librerie fagiche di frammenti di cDNA codificanti per frammenti ORF, e che quindi potrebbero corrispondere a potenziali epitopi. Questo tipo di librerie contengono, potenzialmente, tutto il repertorio ORF di una cellula o di un tessuto e possono quindi essere utilizzate nello studio di malattie autoimmuni al fine di identificare nuovi epitopi coinvolti nella risposta immunitaria, di fare un confronto tra lo stato patologico e quello sano o tra diverse condizioni patologiche. Abbiamo quindi messo a punto un complesso protocollo che prevede: la normalizzazione del cDNA, la sua frammentazione per ottenere peptidi di dimensioni opportune, e l'arricchimento in frammenti realmente codificanti. Con questo sistema abbiamo realizzato una libreria di epitopi a partire da mRNA di cervello umano.
XXIV Ciclo
1984
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Chen, Yun-Liang. "Cloning and expression of sperm antigens from a human testis lambda (#lambda#) gt11 cDNA library." Thesis, Queen Mary, University of London, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.325529.

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Collins-De, Peyer Laurence. "Screening of a rat thymus and a human hippocampus cDNA library for a novel fyn-related oncogene." Thesis, Hong Kong : University of Hong Kong, 1999. http://sunzi.lib.hku.hk/hkuto/record.jsp?B21253870.

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Green, Melanie Leslie Dawn. "The identification of novel autoantigens by means of serological screening of a cDNA expression library constructed from multiple sclerosis brain tissues." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape8/PQDD_0018/MQ54892.pdf.

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Liang, Binhua. "Molecular cloning and sequence analysis of a human brain cDNA of an Alzheimer amyloid precursor (APP) interacting protein." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape7/PQDD_0004/MQ45086.pdf.

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Yamakawa, Tatsuya. "Screening of human cDNA library reveals two differentiation-related genes, HHEX and HLX, as promoters of early phase reprogramming toward pluripotency." Kyoto University, 2016. http://hdl.handle.net/2433/217142.

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Tseng, Huan-Yi, and 曾煥怡. "Isolation and Characterization of GP-83 encoding cDNA from cDNA library of human epididymis." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/74242961586346401682.

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碩士
國防醫學院
解剖學研究所
83
Mammalian sperm are immature and incapable of fertilization when they just emerge from the testis. During the subsequent passage in epididymis, sperm interact with epididymal secretions, and acquire forward motility and ability to recognize and penetrate zona pellucida of eggs. Several genes of sperm maturation-related proteins, such as PH-30, EAP-1, HE 2 and 3 have been isolated and sequenced. DNA sequence of these molecules reveal proteinase- and disintegrin-encoding domains which are the same as those encoded in the genes of a variety of snake venoms. Our previous studies identified a sperm maturation-related glycoproteins, GP-83, in human epididymis. It was secreted from corpus of epididymis will associated in sperm surface. In order to analyze DNA sequence and predict the biological significance, GP-83 encoding cDNA was isolated from cDNA library of human epididymis by immunoscreening using GP-83 specific antiserum. Three positive clones containing inserts of 1.8 Kb, 1.5 kb and 4.0 kb were isola ted. Partial sequences of these inserts, up to 300-400 bp from 3' end, did not reveal any significant homology to known genes in the GenBank. In cDNA library of snake venomous gland, however, an ancrod-like sequence was identified. In conclusion, GP-83 may be a human epididymis-specific protein containing disintegrin-like domain.
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"Identification of human cytosolic malate dehydrogenase by large scale human heart cDNA library sequencing." Chinese University of Hong Kong, 1995. http://library.cuhk.edu.hk/record=b5895590.

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by Agnes, Lo Shuk Yee.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1995.
Includes bibliographical references (leaves [27-33] (2nd gp.)).
Chapter PART 1: --- Human Heart cDNA Library Sequencing
Chapter A) --- Introduction of human heart cDNA library sequencing
Chapter A.1 --- Human genome project
Chapter A.2 --- The aim of human genome project
Chapter A.3 --- Automatic sequencing
Chapter A.4 --- Cycle sequencing reaction
Chapter A.5 --- Human heart cDNA library sequencing project
Chapter B) --- Methods and materials
Chapter (I) --- Preparation of plating bacterial-Y1090
Chapter (II) --- Plating the bacteriophage with blue-white visual selection
Chapter (III) --- Amplification of bacteriophage cDNA clones by PCR
Chapter (IV) --- Purification and quantitation of PCR products
Chapter (V) --- Cycle DNA sequencing of PCR products
Chapter (VI) --- Casting the sequencing gel
Chapter (VII) --- Sequencing by Pharmacia LKB A.L.F. DNA Sequencer
Chapter (VIII) --- Editing and saving the DNA sequence
Chapter (IX) --- Sending the DNA sequence to Genbank by E-mail
Chapter (X) --- Usage of the Genbank database
Chapter C) --- Results
Chapter D) --- Discussions
Chapter D.1 --- Application of human genomic project
Chapter D.2 --- Interpretation of the sequencing results
Chapter D.3 --- Quality of cDNA libraries and representation of mRNA population
Chapter D.4 --- "Gene expression profile in three different organs-heart, brain and liver"
Chapter D.5 --- Population study of the cDNA library
Chapter D.6 --- Isolation of a large number of novel genes by substraction cDNA library
Chapter D.7 --- Screening method to find out the complete coding sequence of interesting genes
Chapter D.8 --- Technical problems encountered and managed
Chapter PART 2: --- Identification of human cytosolic malate dehydrogenase by large scale human heart cDNA library sequencing
Chapter CHAPTER 1: --- Introduction of malate dehydrogenase
Chapter 1.1 --- Malate dehydrogenase--Kreb's cycle enzyme
Chapter 1.2 --- Two stereospecific forms of dehydrogenase
Chapter 1.3 --- NAD-binding domain
Chapter 1.4 --- The active site
Chapter 1.5 --- Comparison of surface properties between cMDH and mMDH
Chapter 1.6 --- N-terminal region and mitochondrial import
Chapter 1.7 --- Subunit-subunit interactions
Chapter 1.8 --- Physiological importance of malate dehydrogenase
Chapter 1.9 --- Secondary structure-total 11 β-strands and 9 α-helixes
Chapter 1.10 --- Objectives of the thesis
Chapter CHAPTER 2: --- Cloning and sequence analysis of human cytosolic malate dehydrogenase (hcMDH)
Chapter 2.1 --- Cloning of human cytosolic malate dehydrogenase (hcMDH)
Chapter 2.1.1 --- Methods and materials
Chapter 2.1.1.1 --- Cloning full length of hcMDH into expression vector pAED4
Chapter 2.1.1.2 --- Preparation of competent cell-JM109 for transformation
Chapter 2.1.1.3 --- Minipreparation of plasmid DNA
Chapter 2.1.1.4 --- Midi-preparation of bacteriophage λDNA by QIAGEN´ёØ
Chapter 2.1.1.5 --- Titration of bacteriophage λ of human adult heart cDNA library
Chapter 2.1.1.6 --- Preparation of soft-agarose lysates
Chapter 2.1.1.7 --- Elution of DNA from agarose gel by GENECLEAN´ёØ
Chapter 2.1.2 --- Results
Chapter 2.1.3 --- Discussions
Chapter 2.2 --- Sequence analysis of human cytosolic malate dehydrogenase (hcMDH)
Chapter 2.2.1 --- Methods and materials: Autoread sequencing
Chapter (I) --- Annealing of primer to double-stranded template
Chapter (II) --- Sequencing
Chapter 2.2.2 --- Results and discussions
Chapter 2.3 --- Amino acids and protein structure analysis of cMDH
Chapter CHAPTER 3 : --- "Protein expression, partial purification and folding experiments of human cytosolic malate dehydrogenase (hcMDH)"
Chapter 3.1 --- Protein expression of hcMDH in E. coli
Chapter 3.1.1 --- Methods and materials
Chapter 3.1.1.1 --- Protein expression induced by IPTG
Chapter 3.1.1.2 --- Isoelectric focusing (IEF)-two dimensional gel electrophoresis
Chapter (I) --- First dimensional electrofocusing
Chapter (II) --- The second dimension SDS-PAGE electrophoresis
Chapter (III) --- Sample preparation
Chapter 3.1.2 --- Results
Chapter 3.1.3 --- Discussions
Chapter 3.1.3.1 --- The properties of expressed protein of hcMDH
Chapter 3.1.3.2 --- T7 expression system
Chapter 3.1.3.3 --- Strong φ 10 promoter
Chapter 3.1.3.4 --- E.coli BL21 host cell
Chapter 3.2 --- Partial purification and folding experiments of hcMDH
Chapter 3.2.1 --- Methods and materials
Chapter 3.2.1.1 --- Partial purification of hcMDH expressed protein
Chapter (I) --- Preparation of supernatant from E.coli crude extract
Chapter (II) --- Ion-exchange column chromatography
Chapter (III) --- Affinity chromatography
Chapter (IV) --- Gel filtration on a Sepharose CL-6B column
Chapter 3.2.1.2 --- Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)
Chapter 3.2.1.3 --- Staining the protein gel by the Coomassie Blue R-250 method
Chapter 3.2.1.4 --- Staining the protein gel by the Silver staining Method
Chapter 3.2.1.5 --- Quantitation of protein by the Bradford Method
Chapter 3.2.1.6 --- Native gel electrophoresis
Chapter 3.2.1.7 --- Malate dehydrogenase MDH enzyme staining method
Chapter 3.2.1.8 --- Malate dehydrogenase MDH enzyme assay
Chapter 3.2.1.9 --- Fast protein liquid chromatography (FPLC)
Chapter 3.2.1.10 --- Protein folding experiment
Chapter 3.2.1.11 --- Eukaryotic expression of hcMDH
Chapter 3.2.2 --- Results
Chapter 3.2.2.1 --- Partial purification by chromatography
Chapter 3.2.2.2 --- Native gel
Chapter 3.2.2.3 --- FPLC
Chapter 3.2.2.4 --- To aid folding of protein by adding NADH
Chapter 3.2.2.5 --- Eukaryotic expression
Chapter 3.2.3 --- Discussions
Chapter 3.2.3.1 --- Purification of malate dehydrogenase MDH
Chapter 3.2.3.2 --- "Methods for visualizing dehydrogenase enzymes, e.g. malate dehydrogenase"
Chapter 3.2.3.3 --- The presence of unfold hcMDH protein in bacteria
Chapter 3.2.3.4 --- Folding of protein by heat shock protein GroE
Chapter 3.2.3.5 --- Eukaryotic expression
Chapter CHAPTER 4: --- Master screening of single base change by PCR-SSCP (Single Strand Conformational Polymorphism)
Chapter 4.1 --- Theory of SSCP
Chapter 4.2 --- Methods and materials
Chapter 4.3 --- Results
Chapter 4.4 --- Discussions
Chapter 4.4.1 --- The procedure of SSCP
Chapter 4.4.2 --- An alternative quick detection method for polymorphism of hcMDH at position 565--by automatic sequencing
Chapter 4.4.3 --- Other detection methods-- RNA-PCR and ddF
Chapter 4.4.4 --- Parameters affecting sensitivity of SSCP
Chapter 4.4.5 --- Application of SSCP
Chapter CHAPTER 5: --- Southern hybridization and In situ hybridization
Chapter 5.1 --- Southern blot analysis of human cytosolic malate dehydrogenase (hcMDH)
Chapter 5.1.1 --- Methods and materials
Chapter (I) --- Transfer genomic DNA to Nylon membrane
Chapter (II) --- Synthesis of radiolabelling cDNA probe
Chapter (III) --- Pre-hybridization and hybridization reaction
Chapter 5.1.2 --- Results
Chapter 5.1.3 --- Discussions
Chapter 5.2 --- In situ hybridization
Chapter 5.2.1 --- Methods and materials
Chapter (I) --- Preparation of Dig labelling probe by random primed labelling
Chapter (II) --- Estimating the yield of Dig-labelled nucleic acids
Chapter (III) --- Denaturation and hybridization of the hcMDH probe with animal tissues
Chapter (IV) --- Color development of the tissue
Chapter 5.2.2 --- Results
Chapter 5.2.3 --- Discussions
Chapter 5.2.3.1 --- Cellular distribution of hcMDH
Chapter 5.2.3.2 --- The principle of in situ hybridization
Chapter 5.2.3.3 --- Specimen preparation
Chapter 5.2.3.4 --- Hybridization conditions
Chapter 5.2.3.5 --- "Ontogeny of MDH in rabbit fetal brain, heart and lung"
Appendixes:
"Appendix I: 531 random cDNA clones from clone no. J950 to K951 in human heart cDNA library sequencing project. The name of clones, accession number, the length of the partial sequence and percentage of match are listed"
Appendix II: The new accession no. of Novel clones in Genbank
"Appendix III: The enzymatic reaction, molecular weigth, specific activity and Michaelis constants of different sources of malate dehydrogenase"
Appendix IV: The full sequence of nucleic acids and amino acids of human cytosolic malate dehydrogenase hcMDH. Accession no. of hcMDH is U20352 in Genbank
Appendix V: Nucleotide sequences of the mouse cMDH gene
Appendix VI: Nucleotide sequences of the mouse mMDH gene
Appendix VII: Structural organization of the mouse cytosolic malate dehydrogenase and its comparison with that of the mouse mitochondrial malate dehydrogenase gene
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9

"Gene expression of adult human heart as revealed by random sequencing of cDNA library." Chinese University of Hong Kong, 1995. http://library.cuhk.edu.hk/record=b5895472.

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by Tsui Kwok-wing.
Thesis (Ph.D.)--Chinese University of Hong Kong, 1995.
Includes bibliographical references (leaves 188-216).
ACKNOWLEDGEMENTS --- p.ii
ABSTRACT --- p.iii
TABLE OF CONTENTS --- p.v
ABBREVIATIONS --- p.ix
Chapter CHAPTER 1 --- INTRODUCTION
Chapter 1.1 --- General introduction --- p.1
Chapter 1.2 --- Human genome project --- p.5
Chapter 1.3 --- Organization of human genome --- p.7
Chapter 1.4 --- Adult human heart cDNA library --- p.9
Chapter 1.5 --- Gene expression in adult human heart --- p.10
Chapter 1.6 --- Polymerase chain reaction --- p.12
Chapter 1.7 --- Purification of PCR products --- p.15
Chapter 1.8 --- Automated DNA sequencing --- p.17
Chapter 1.9 --- Sequence analysis by electronic mail server --- p.21
Chapter 1.10 --- Effects of agar and agarose on Vent´ёØ and Taq DNA polymerases --- p.23
Chapter 1.11 --- Transcription factors and zinc finger proteins --- p.25
Chapter 1.12 --- LIM domain --- p.28
Chapter 1.13 --- Cysteine-rich intestinal protein --- p.30
Chapter CHAPTER 2 --- MATERIALS AND METHODS
Chapter 2.1 --- Plating out the adult human heart cDNA library --- p.32
Chapter 2.2 --- Amplification by polymerase chain reaction --- p.33
Chapter 2.3 --- Purification of the PCR products by Millipore filters --- p.35
Chapter 2.4 --- Elimination of the purification of the PCR products before sequencing --- p.36
Chapter 2.5 --- Cycle sequencing --- p.37
Chapter 2.6 --- Unicycle sequencing --- p.38
Chapter 2.7 --- Sequencing by T7 polymerase --- p.39
Chapter 2.8 --- Gel electrophoresis in the automated A.L.F. sequencer --- p.41
Chapter 2.9 --- Sequence analysis by commercially available softwares --- p.42
Chapter 2.10 --- Sequence analysis through electronic mail server --- p.44
Chapter 2.11 --- Database for storing the result of each clone --- p.46
Chapter 2.12 --- Effects of agar and agarose on Vent´ёØ and Taq DNA polymerase --- p.47
Chapter 2.13 --- Mini-preparation of plasmid DNA --- p.50
Chapter 2.14 --- Large scale preparation of plasmid DNA --- p.51
Chapter 2.15 --- Cloning the human cysteine rich heart protein (hCRHP) into the pAED4 vector --- p.53
Chapter 2.16 --- Expression of hCRHP in E coli --- p.56
Chapter 2.17 --- Northern hybridization --- p.58
Chapter 2.18 --- Partial protein sequencing of hCRHP --- p.59
Chapter CHAPTER 3 --- RESULTS
Chapter 3.1 --- The sequencing results of adult human heart cDNA clones --- p.60
Chapter 3.2 --- Accuracy of sequencing results --- p.63
Chapter 3.3 --- Catalogues of genes expressed in the adult human heart --- p.65
Chapter 3.4 --- Effects of agar and agarose on Vent´ёØ and Taq DNA polymerases --- p.94
Chapter 3.5 --- Elimination of the purification of the PCR products before sequencing --- p.102
Chapter 3.6 --- Sequence analysis of hCRHP --- p.104
Chapter 3.7 --- Northern hybridization of hCRHP --- p.109
Chapter 3.8 --- Expression of hCRHP in E. coli --- p.112
Chapter CHAPTER 4 --- DISCUSSION
Chapter 4.1 --- Random sequencing of adult human heart cDNA clones --- p.118
Chapter 4.2 --- Catalogues of genes expressed in the adult human heart --- p.130
Chapter 4.3 --- Gene expression in the adult human heart --- p.137
Chapter 4.4 --- Importance of nonhuman matches --- p.170
Chapter 4.5 --- Effects of agar and agarose on Vent´ёØ and Taq DNA polymerases --- p.177
Chapter 4.6 --- Elimination of the purification of the PCR products before sequencing --- p.180
Chapter 4.7 --- The possible role of CRIP and hCRHP --- p.184
Chapter 4.8 --- Future prospect --- p.186
REFERENCE --- p.188
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LIN, YI-LING, and 林宜玲. "Analysis of ▫ 36A, anintegrated HBV DNA and its cellular flanking sequences clonod from a human hepatocellular carcinoma cell line HCC36, and construction of a cDNA library from HCC 36." Thesis, 1987. http://ndltd.ncl.edu.tw/handle/34740816730930970350.

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Books on the topic "Human brain cDNA library"

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Cao, Chenglong. Immunological screening of a rat brain cDNA library for genes encoding potential novel glutamate receptors. Ottawa: National Library of Canada, 1993.

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The river that flows uphill: A journey from the Big Bang to the Big Brain. San Francisco: Sierra Club Books, 1986.

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The river that flows uphill: A journey from the Big Bang to the Big Brain. New York: Macmillan, 1986.

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King, Stephen. Mertvai︠a︡ zona. Moskva: AST, 1997.

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Stephen, King. The dead zone. London: Futura, 1980.

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King, Stephen. La zona muerta. 4th ed. Barcelona, Spain: Plaza & Janés, 1998.

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Stephen, King. The dead zone. London: Warner Books, 1992.

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King, Stephen. The dead zone. Boston: G.K. Hall, 1993.

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King, Stephen. Nekre zone. Athens: Bell, 2001.

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King, Stephen. The Dead Zone. New York: Penguin USA, Inc., 2009.

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Book chapters on the topic "Human brain cDNA library"

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Mukherjee, Arkajyoti, Ritik Srivastava, Vansh Bhatia, Utkarsh, and Suneeta Mohanty. "Stimuli Effect of the Human Brain Using EEG SPM Dataset." In Intelligent Systems Reference Library, 213–26. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-37551-5_14.

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Fukuoka, Shin-Ichi, and Katsumi Shibata. "Characterization and Functional Expression of the Cdna Encoding Human Brain Quinolinate Phosphoribosyltransferase." In Advances in Experimental Medicine and Biology, 611–14. Boston, MA: Springer US, 1999. http://dx.doi.org/10.1007/978-1-4615-4709-9_76.

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Grübler, Gerd, and Elisabeth Hildt. "On Human–Computer Interaction in Brain–Computer Interfaces." In The International Library of Ethics, Law and Technology, 183–91. Dordrecht: Springer Netherlands, 2014. http://dx.doi.org/10.1007/978-94-017-8996-7_15.

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Liew, C. C., D. M. Hwang, R. X. Wang, S. H. Ng, A. Dempsey, D. H. Y. Wen, H. Ma, et al. "Construction of a human heart cDNA library and identification of cardiovascular based genes (CVBest)." In Novel Methods in Molecular and Cellular Biochemistry of Muscle, 81–87. Boston, MA: Springer US, 1997. http://dx.doi.org/10.1007/978-1-4615-6353-2_8.

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Salehi, Zahra, Susan Ramos, Gary Pearson, Mira Jung, Anatoly Dritschilo, and Francis G. Kern. "Construction of a Unidirectional cDNA Library from a Radioresistant Laryngeal Squamous Cell Carcinoma Cell Line in an Epstein Barr Virus Shuttle Vector." In Neoplastic Transformation in Human Cell Culture, 377–86. Totowa, NJ: Humana Press, 1991. http://dx.doi.org/10.1007/978-1-4612-0411-4_38.

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Pham, Dung Ngoc. "Profiling General-Purpose Fast Multipole Method (FMM) Using Human Head Topology." In Brain and Human Body Modeling 2020, 347–81. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-45623-8_21.

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AbstractIn this study, we characterize the performance of the fast multipole method (FMM) in solving the Laplace and Helmholtz equations. We use the FMM library developed by the group of Dr. L. Greengard. This version of the FMM algorithm is multilayer with no priori limit on the number of levels of the FMM tree, although, after about thirty levels, there may be floating point issues. A collection of high-resolution human head models is used as test objects. We perform a detailed analysis of the runtime and memory consumption of the FMM in a wide range of frequencies, problem sizes, and precisions required. Although we focus on two-manifold test cases, the results are generalizable to other topologies as well. The tests are conducted on both Windows and Linux platforms. The results obtained in this study can serve as a general benchmark for the performance of FMM. It can also be employed to pre-estimate the efficiency of numerical modeling methods (e.g., the boundary element method) accelerated by FMM.
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Wang, Ruoxiang, Eva Cukerman, Baosheng Chen, and Choong-Chin Liew. "Differential Screening and Megasequencing of Human Heart cDNA Library: A Search for Genes Associated with Heart Failure." In Developments in Cardiovascular Medicine, 67–77. Boston, MA: Springer US, 1995. http://dx.doi.org/10.1007/978-1-4613-1237-6_6.

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PARK, M. S., P. E. PARDINGTON, and D. J. CHEN. "FUNCTIONAL COMPLEMENTATION OF A DNA DOUBLE STRAND REPAIR DEFICIENT CHINESE HAMSTER MUTANT WITH HUMAN cDNA LIBRARY." In Radiation Research: A Twentieth-century Perspective, 329. Elsevier, 1991. http://dx.doi.org/10.1016/b978-0-12-168561-4.51102-1.

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Horellou, Philippe, Lionel Marlier, Alain Privat, François Darchen, Daniel Scherman, Jean-Pierre Henry, and Jacques Mallet. "Chapter 3 Exogeneous expression of L-dopa and dopamine in various cell lines following transfer of rat and human tyrosine hydroxylase cDNA: grafting in an animal model of Parkinson's disease." In Progress in Brain Research, 23–32. Elsevier, 1990. http://dx.doi.org/10.1016/s0079-6123(08)62586-8.

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Ruzic, Fjodor. "Information-Communications Systems Convergence Paradigm." In Human Computer Interaction, 2324–40. IGI Global, 2009. http://dx.doi.org/10.4018/978-1-87828-991-9.ch155.

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This chapter is on cultural aspects of information- communications systems embedded into new media environment and invisible e-technologies, and on a new age of social responsibility for information technology professionals. Besides the key issues in information technology development that create smart environment and ambient intelligence, the chapter also discusses digital e-culture and the new media role in cultural heritage. From the viewpoint of information technology, the current information-communications systems converge with media. This convergence is about tools-services-content triangle. Thus, we are confronted with a new form of media mostly presented with the term digital, reshaping not only media industry but also a cultural milieu of an entire nation on a regional and global basis. The discussion follows on the World Library idea that is rebuilding with new form of World Memory (World Brain), the shift from visible culture domination to the domination of invisible culture in the world of e-technologies predominance. From this scenario, information technology professionals coping with information systems projects, e-services development, and e-content design have more cultural responsibility than in the past when they worked within closer and inner cultural horizons and when their misuse of technologies had no influence on culture as a whole.
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Conference papers on the topic "Human brain cDNA library"

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Edgington, T. S., J. H. Morrissey, and H. Fakhrai. "MOLECULAR CLONING OF HUMAN TISSUE FACTOR cDNA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643740.

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Tissue factor (TF), the cell-surface receptor and allo-steric activator for factor Vll/VIIa, is important in hemostasis andinflammation. The TF apoprotein was purifiedfrom human brain using factor Vll-affinitychromatography and SDS gel electrophoresis,and was found to consist of a 47 kDa heavy chain plus a 12.5 kDa light chain. Approximately one-third of the heavy chain amino acid sequence was determined for four regions by microsequencing the intact protein and peptides derived from V8-protease digestion. A λgtll cDNA library, made from mRNA derived from the human fibroblastic cell line WI38,was screened with (a) affinity-purified rabbit antibodies to human tissue factor, and (b) a 45-mer oligonucleotide probe based on TF heavy chain amino acid sequence. Five overlapping cDNA clones were identifiedand sequenced which confirmed all four partial TF amino acid sequences. Together these clones span the entire heavy chain coding sequence as well as 5" and 3" nontranslated regions. The N-terminusof the TF heavy chain is preceded by an unusually long signal peptide which appears to be cleaved at alternative sites two amino acids apart. This results in two variants of TF heavy chains which differ slightly in length and amino-terminal sequence.The deduced protein sequence shows no major homology to known protein sequences. However,a relatively uncommon tripeptide sequence, Trp-Lys-Ser (WKS), appears three times in the TF heavy chain. This tripeptidesequence also occurs in HMW kininogen, factor VIII,von Willebrand"s factor andant ithrombin-III. Limited sequence similarity is observed in flanking sequences,andthis may indicate a possible functional domain for the recognition of members ofthe vitamin K-dependent serine protease famil.Supported by NIH grants HL-16411 andCA-41085.
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Morrissey, J. H., D. S. Fair, and T. S. Edgington. "STRUCTURE AND PROPERTIES OF THE HUMAN TISSUE FACTOR APOPROTEIN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643738.

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Tissue factor (TF), an integral membrane glycoprotein, is an initiating molecule for the coagulation protease cascade. TF must reside in a phospholipid membrane for optimal activity where it functions as the receptor and essential allosteric activator for factor Vll/VIIa.TF apoprotein was purified from human brain and placenta using factor Vll-affinity chromatography or immunoaffinity chromatography with a mouse anti-TF monoclonal antibody. Both methods resulted in a homogeneous preparation consisting of a highly glycosylated 47 kDa heavy chain and a 12.5 kDa light chain.Removal of asparagine-linked carbohydrate chains with glycopeptidase F reduced the apparentmolecular weight of the heavy chain to 37 kDa but hadno effect on the mobility of the light chain in SDS gel electrophoresis. Electrophoretic analysis of theintact protein with and without reduction indicates that the light chain is disulfide-linked to the heavychain in about half of the TF molecules and is notessential for function.The majority of polyclonal andtwenty- nine monoclonal antibodies against purified TF strongly inhibit coagulation and in all cases aredirected against epitopes on the heavy chain alone. Functional regions of the TF heavy chain have been investigated using a library of twenty-nine monoclonalantibodies and a series of overlapping, synthetic oligopeptides based on sequence information obtained from cloning the cDNA for TF. Supported by NIH grantsHL-16411 and CA-41085.
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Bjorklid, E., T. Johansen, U. R. Pendurthi, L. V. M. Rao, B. Warn-Cramer, and S. T. Rapaport. "HUMAN cDNA CLONES FOR THROMBOPLASTIN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643735.

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A rabbit polyclonal antibody which was monospecific for thromboplastin (TP) apoprotein from human brain according to a number of criteria was used to screen two human placenta cDNA libraries in the expression vector λgt11, one randomly primed and one oligo-dT primed, by the method of Young and Davis. 23 positive clones expressing TP related antigen were isolated and plaque purified. DNA from the different clones was isolated and the TPcDNA inserts released by EcoRl digestion. The inserts could be classified into11 size classes ranging from approx. 300-1100 base pairs.The largest insert (1100 bp) was subcloned intothe plasmid vector pGEM-1. When the nick-translated plasmid (pTP4-l) was used as a probe to screen the phage clones by slot blot hybridization all the 23clones hybridized to the 1100 bp insert. A λgt11TP4 lysogen expressed β-galactosidase- TP4 fusion peptide upon IPTG induction as shown by immunobinging studied using two different antibodies to TP apoprotein: the rabbit antibody originally used to screen the libraries and an antibody raised in goat against human brain TP purified by affinity chromatography on a Factor VII-antiVII-agarose column.
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Beeler, D., L. Fritze, G. Soff, R. Jackman, and R. Rosenberg. "HUMAN THROMBOMODULIN cDNA:SEQUENCE AND TRANSLATED STRUCTURE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643967.

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A 750 bp bovine Thrombomodulin (TM) cDNA fragment was used as an hybridization probe to screen an oligo-dT primed Lambda gtll. cDNA library prepared from human umbilical vein endothelial cell mRNA. A 2.4 kb positive human clone was isolated which showed an 80% nucleotide sequence homology with bovine TM cDNA. This clone and a 550 bp fragment from its 5' end were used to further screen the oligo-dT primed library as well as randomly primed library prepared from the same mRNA. The cDNA clones obtained allow us to describe the overall structure of human TM and reveal that it is extremely similar to the structure of bovine TM, especially as the bovine TM is organized like the receptor for low density lipoprotein (LDL R). Both TM and LDL R exhibit short cytoplasmic C-terminal tails which are either neutral or negatively charged. Other coated pit receptors such as the insulin receptor or the epidermal growth factor (EGF) receptor have very large cytoplasmic regions with a complex tyrosine kinase segment as well as multiple sites for phosphorylation. Both TM and LDL R possess a transmembrane region and an immediately adjacent extracellular serine/threonine rich region which in LDL R has been shown to bear 0-1inked sugars. Both TM and LDL R contain a more distal area of cysteine rich repeats, first noted in the EGF precursor and termed EGF type B. However, the TM EGF type B repeats appear to have been duplicated in TM resulting in their being 6 of them rather than the 3 found in LDL R. The N-terminal half of LDL R is thought to contain the ligand binding region of the receptor and is constructed from multiple cysteine rich repeats similar to those of Complement factor C9. The structure of this region of TM is quite different from that of LDL R, possessing few cysteines. We suspect that protein C and/or thrombin may bind to this unique domain of TM.
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Dahlbäck, B., and A. LundWall. "ISOLATION AND CHARACTERIZATION OF cDNA CLONES FOR HUMAN FACTOR V." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643886.

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Coagulation factor V is a single chain, 330 kDa glycoprotein functioning as a cofactor to factor Xa in the activation of prothrombin. Thrombin cleaves factor V into four major fragments, out of which the N-terminal (105kDa) and the C-terminal (71-74kDa) fragments together constitute the active factor V species. To isolate cDNA clones a λ-gt 11 liver library was screened with a polyclonal, monospecific antiserum against human factor V. Four positive clones (two "weak", Aland A2 and two "strong", A3 and A4) were identified and isolated. Al(0.7kb), A2 (1.25kb) and A4 (0.85kb) reacted strongly with an antiserum against the 105 kDa, N-terminal fragment (heavy chain of factor Va), whereas A3 (1.25kb) gave the best signal with an antiserum against the 71-74 kDa, C-terminal fragment (light chain of factor Va). A1 hybridized with A2 and A4, whereas A2 only hybridized with Al. A3, which did not hybridize to any of the other clones, was used to rescreen the library and 9 positive clones (Bl-9) were isolated. B9 (3kb) coded for the entire C-terminal factor V fragment and the 3' noncoding sequence. B8 (1.8kb) partially overlapped B9 but extented the 5' sequence with 0.8kb. In a third screening round Al was used in combination with B8 and a 1.1 kb clone (CIO) was identified which hybridized to both. C10 did not hybridize with A2. The following overlapping cDNA clones can be orderedfrom the 5´end: A2-A1-C10-B8-B9 and together they cover 6 kb of coding sequence
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Sumi, Y., Y. Nakamura, M. Sakai, M. Muramatsu, and N. Aoki. "STRUCTURE OF HUMAN α2-PLASMIN INHIBITOR DEDUCED FROM THAT OF cDNA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644371.

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The complete amino acid sequence of α2-plasmin inhibitor (α2PI) was determined by cDNA cloning. A Agt 10 cDNA library was prepared from poly(A)+mRNA isolated from cultured human liver cells. The labeled oligonucleotides, corresponding to the reported partial amino acid sequences of α2PI, were used as probes to screen the library. One of the positive clones was subcloned into plasmid pUC8. A 2.2 kilobase cDNA clone thus isolated contains a region coding for a portion of a leader sequence, the mature protein, a stop codon (TGA), a 3' noncoding region (733 nucleotides), and a poly(A)tail (37 nucleotides). The amino acid sequence deduced from the cDNA is composed of 452 amino acids starting with an amino-terminal sequence of Asn-Gln-Glu-Gln and ending with a carboxyl-terminal sequence of Gly-Ser-Pro-Lys. The sequence shows approximately 30% homology with those of other plasma serine protease inhibitors. However, α2PI extends 50-52 amino acids beyond the carboxyl-terminal ends of the other inhibitors. This 50-52 carboxyl-terminal amino acid sequence is therefore specific to α2PI, and contains the sequence that is exactly the same as that of the peptide containing the plasminogen binding site. There are three lysine residues possibly involved in the binding to plasminogen in this region. From the homology with the other inhibitors, the inhibitor's reactive-site peptide bond was suggested to be Met-Ser and the same as that of ai-antitrypsin. The Met residue is located at the 362 position from the amino-terminal end.
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Ploos van Amstel, J. K., A. L. van der Zanden, E. Bakker, P. H. Reitsma, and R. M. Bertina. "INDEPENDENT ISOLATION OF HUMAN PROTEIN S cDNA AND THE ASSIGNMENT OF THE GENE TO CHROMOSOME 3." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644638.

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Protein S is a vitamin K-dependent glycoprotein, that serves as a cofactor of activated protein C. A hereditary deficiency in protein S is associated with an increased risk for the development of venous thrombosis. The deficiency is inherited as an autosomal dominant trait. We isolated a cDNA coding for protein S and assigned its gene to chromosome 3.A human liver cDNA library in phage xgtll (complexity 1.2 × 106 , D. Stafford, Chapel Hill) was screened by using immuno-purifiedpolyclonal anti-protein S IgG as a probe. Approximately 1.5 x 10 recombinants of the amplified library were screened. Out of eighteen positive clones one clone was found, after nucleotide sequence analysis, to code for a peptide with a high degree of homology with the carboxy terminal region of the already published bovine protein S. This clone pSP84 (450 bp) was used as a probe to screen a human liver cDNA library in plasmid pUC9. From this library we isolated several positive clones. Clone pSUL5 contained the largest insert (2200 base pairs). Dideoxy sequencing revealed that it codes for 330 amino acids of the carboxy terminal part of protein S. Furthermore, it contained a 1200 base pairs 3' untranslated region. The predicted amino acid sequence did not differ from the published sequence of human protein S, although at the nucleotide level some differences could be detected.Clone pSUL5 was used to localize the protein S gene to its chromosome. The assignment was done by hybridization to Pst I digested DNA from human-hamster c.q. human-mouse somatic cell hybrids. In this way we got strong indication that the protein S gene is located on chromosome 3.
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Koide, T. "CHARACTERIZATION OF THE GENE FOR HUMAN HISTIDINE-RICH GLYCOPROTEIN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643599.

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Human histidine-rich glycoprotein (HRG) is a single-chain glycoprotein in plasma which is considered to modulate a coagulation and fibrinolysis system with the ability to bind to heparin, plasminogen, fibrinogen, thrombospondin, etc. Recently we have elucidated the primary structure of HRG by determining the nucleotide sequence of its cDNA, and showed that HRG is composed of several different types of internal repeats, each one of which shows considerable homology with the functional and/or structural domains of other proteins including high molecular weight kininogen, antithrombin III, cystatins, and proline-rich protein and peptide. Thus, the multifunctional property of HRG was suggested to be due to its multi-domain structure. In the present studies, a human genomic DNA library, cloned in the bacteriophage vector Charon 4A, was screened for HRG gene using a full-length cDNA coding for human IMI as a probe. A total of 7 clones were isolated from 6 × 105 phage and each was plaque purified. The entire HRG gene is represented in 3 genomic inserts with overlapping sequences that carry human DNA spanning 30 kb. Overlapping gene fragments were subcloned into pUC9 and characterized by Southern blot hybridization using 5’ and 3’ end probes isolated from human HRG cDNA and by DNA sequencing. These studies have shown that the gene for human HRG spans about 9 kb and consists of at least 5 exons and 4 introns. The putative histidine-rich region consisted of 12 tandemly repeated sequences of a 5 amino acid segment and 2 proline-rich regions contiguous to it are likely to be involved within one exon.
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Bosma, P. J., E. A. van den Berg, and T. Kooistra. "ISOLATION OF THE GENE CODING FOR HUMAN PLASMINOGEN ACTIVATOR INHIBITOR TYPE 1 (PAI-1)." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644440.

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A human placenta genomic DNA cosmid library was screened for the presence of the PAI-1 gene using a cDNA probe coding for PAI-1. Two overlapping recombinant cosmids were obtained that contain human DNA spanning 55 kb. The cosmids were mapped using 3' and 5' end probes isolated from an almost full-length cDNA clone of 2.5 kb. The two cosmids were found to contain the entire structural PAI-1 gene (approximately 15 kb) and also included 25 kb 5' flanking sequences. The transcription initiation site was identified by SI nuclease protection experiments and the promotor region was sequenced. Further experiments will be directed at characterizing the regulatory elements of the PAI-1 gene.In order to determine the chromosomal localization of the PAI-1 gene we have hybridized our genomic clones in situ to metaphase chromosomes of a human blood cell culture. Preliminary experiments show a specific hybridization signal which will enable us to sublocalize the chromosomal position of the PAI-1 gene.
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van den Berg, E. A., E. Sprengers, M. Jaye, W. Burgess, and V. W. M. van Hinsbergh. "REGULATION OF PLASMINOGEN ACTIVATOR INHIBITOR-1 mRNA IN HUMAN ENDOTHELIAL CELLS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1642856.

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Cultured human endothelial cells (HEC) increase their production of plasminogen activator inhibitor (PAI-1) upon stimulation with endotoxin and IL-1, agents that are known to cause an increase in PAI-1 levels in vivo. In order to study the regulation of PAI-1 synthesis at the mRNA level, we isolated a cDNA clone for the human PAI-1 gene from an endothelial expression cDNA library in λ gt 11 by screening with a PAI-1 specific antibody. Three positive cross-hybridizing clones were isolated. The longest insert (1500 bp) was partially sequenced (1000 bp). The sequence was identical to the PAI-1 sequence recently reported by others. The identity of the cDNA clone was further confirmed by comparison with part of the amino acid sequence of PAI-1. For that purpose t-PA-PAI-1 complex was purified from HEC conditioned medium by immunoadsorption to anti-t-PA IgG, and a suitable peptide was sequenced after comparison of the HPLC elution profiles of CNBr digests of t-PA and t-PA-PAI-1 complex. The amino acid sequence (M)FRQFQADFT completely matches the sequence predicted from the cDNA sequence.By hybridization of the cDNA probe to Northern blots of total cellular RNA from human umbilical vein and artery EC (HUVEC, HUAEC), two transcripts of 2.3 and 3 kb were found. Primary HUAEC, incubated for 18 hours in growth medium, produced considerable although variable levels of PAI-1 activity and contained PAI-1 mRNA levels comparable to those found in subcultured HUAEC. When subcultured HUEC were incubated for 6 h with endotoxin, IL-1 or TNF, a 2-fold increase in PAI-1 mRNA was found with each of these mediators. Stimulation of the cells in the presence of cycloheximide resulted in a further increase of the 3 kb PAI-1 transcript. The 3’ end of this transcript contains a 75 bp AT-rich sequence. Similar 3’ AT-rich sequences have been found in mRNA’s for a number of inflammatory mediators and cellular oncogenes, and in some cases it has been shown that removal of the sequence increased mRNA stability. The influence of cyclohex-imid on the larger PAI-1 transcript might be explained by inhibition of synthesis of a specific nuclease that controls the level of mRNA’s harbouring such an AT rich sequence.
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Reports on the topic "Human brain cDNA library"

1

Sikela, J. M. cDNA expression map of the human genome: Methods development and applications using brain cDNAs. Office of Scientific and Technical Information (OSTI), January 1991. http://dx.doi.org/10.2172/5605006.

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Sikela, J. M. cDNA expression map of the human genome: Methods development and applications using brain cDNAs. Progress report, October 15, 1991--March 14, 1992. Office of Scientific and Technical Information (OSTI), December 1991. http://dx.doi.org/10.2172/10112745.

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