Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire
Identifieur interne : 004810 ( Main/Merge ); précédent : 004809; suivant : 004811Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire
Auteurs : Michael J. Campbell [États-Unis] ; Andrew D. Zelenetz [États-Unis] ; Shoshana Levy [États-Unis] ; Ronald Levy [États-Unis]Source :
- Molecular Immunology [ 0161-5890 ] ; 1992.
English descriptors
- Teeft :
- Acad, Amplification, Amplification products, Autoimmune diseases, Cell clone, Cell malignancies, Clone, Consensus sequences, Gene, Gene expression, Gene families, Gene family usage, Gene usage, Germline, Germline gene, Germline genes, Heavy chain, Homology, Human germline, Human immunoglobulin, Immunoglobulin, Intron sequences, Leader primers, Leader region primers, Leader sequences, Natn, Nucleotide, Nucleotide sequence, Nucleotide sequences, Plasmid, Polymerase, Polymerase chain reaction, Primer, Primer sequences, Proc, Sanz, Schroeder, Sequencing, Variable region genes.
Abstract
Abstract: We have designed a set of six, non-degenerate oligonucleotide primers, corresponding to the 5' leader regions of each of the six human VH gene families. A general strategy for family specific polymerase chain reaction amplification is described using these primers and a conserved 3' primer corresponding to frame work 3, JH, or constant region. This strategy was used to isolate and sequence novel human germline VH genes belonging to the VH2 and VH4 families. Under certain conditions, chimeric VH sequences were created by a “jumping polymerase chain reaction”, combining DNA segments from different germline genes, but this could be avoided by limiting the number of amplification cycles. PCR amplification with these family specific primers will facilitate studies of the repertoire of germline VH genes as well as studies on VH gene usage in normal and aberrant (B cell malignancies, autoimmune diseases, etc.) B cell populations.
Url:
DOI: 10.1016/0161-5890(92)90100-C
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Zelenetz</name></author><author><name sortKey="Levy, Shoshana" sort="Levy, Shoshana" uniqKey="Levy S" first="Shoshana" last="Levy">Shoshana Levy</name></author><author><name sortKey="Levy, Ronald" sort="Levy, Ronald" uniqKey="Levy R" first="Ronald" last="Levy">Ronald Levy</name></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:91039A53C4CF7978D32AED252C61DAB0833E3462</idno><date when="1992" year="1992">1992</date><idno type="doi">10.1016/0161-5890(92)90100-C</idno><idno type="url">https://api.istex.fr/ark:/67375/6H6-3K50034T-5/fulltext.pdf</idno><idno type="wicri:Area/Istex/Corpus">002098</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">002098</idno><idno type="wicri:Area/Istex/Curation">002098</idno><idno type="wicri:Area/Istex/Checkpoint">001D20</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Checkpoint">001D20</idno><idno type="wicri:doubleKey">0161-5890:1992:Campbell M:use:of:family</idno><idno type="wicri:Area/Main/Merge">004810</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Use of family specific leader region primers for PCR amplification of the human heavy chain variable region gene repertoire</title><author><name sortKey="Campbell, Michael J" sort="Campbell, Michael J" uniqKey="Campbell M" first="Michael J." last="Campbell">Michael J. Campbell</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Stanford University School of Medicine, Department of Medicine, Division of Oncology, M207, Stanford, CA 94305-5306</wicri:regionArea><wicri:noRegion>CA 94305-5306</wicri:noRegion></affiliation></author><author><name sortKey="Zelenetz, Andrew D" sort="Zelenetz, Andrew D" uniqKey="Zelenetz A" first="Andrew D." last="Zelenetz">Andrew D. Zelenetz</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Stanford University School of Medicine, Department of Medicine, Division of Oncology, M207, Stanford, CA 94305-5306</wicri:regionArea><wicri:noRegion>CA 94305-5306</wicri:noRegion></affiliation></author><author><name sortKey="Levy, Shoshana" sort="Levy, Shoshana" uniqKey="Levy S" first="Shoshana" last="Levy">Shoshana Levy</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Stanford University School of Medicine, Department of Medicine, Division of Oncology, M207, Stanford, CA 94305-5306</wicri:regionArea><wicri:noRegion>CA 94305-5306</wicri:noRegion></affiliation></author><author><name sortKey="Levy, Ronald" sort="Levy, Ronald" uniqKey="Levy R" first="Ronald" last="Levy">Ronald Levy</name><affiliation wicri:level="1"><country xml:lang="fr">États-Unis</country><wicri:regionArea>Stanford University School of Medicine, Department of Medicine, Division of Oncology, M207, Stanford, CA 94305-5306</wicri:regionArea><wicri:noRegion>CA 94305-5306</wicri:noRegion></affiliation></author></analytic><monogr></monogr><series><title level="j">Molecular Immunology</title><title level="j" type="abbrev">MIMM</title><idno type="ISSN">0161-5890</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1992">1992</date><biblScope unit="volume">29</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="193">193</biblScope><biblScope unit="page" to="203">203</biblScope></imprint><idno type="ISSN">0161-5890</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0161-5890</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Acad</term><term>Amplification</term><term>Amplification products</term><term>Autoimmune diseases</term><term>Cell clone</term><term>Cell malignancies</term><term>Clone</term><term>Consensus sequences</term><term>Gene</term><term>Gene expression</term><term>Gene families</term><term>Gene family usage</term><term>Gene usage</term><term>Germline</term><term>Germline gene</term><term>Germline genes</term><term>Heavy chain</term><term>Homology</term><term>Human germline</term><term>Human immunoglobulin</term><term>Immunoglobulin</term><term>Intron sequences</term><term>Leader primers</term><term>Leader region primers</term><term>Leader sequences</term><term>Natn</term><term>Nucleotide</term><term>Nucleotide sequence</term><term>Nucleotide sequences</term><term>Plasmid</term><term>Polymerase</term><term>Polymerase chain reaction</term><term>Primer</term><term>Primer sequences</term><term>Proc</term><term>Sanz</term><term>Schroeder</term><term>Sequencing</term><term>Variable region genes</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: We have designed a set of six, non-degenerate oligonucleotide primers, corresponding to the 5' leader regions of each of the six human VH gene families. A general strategy for family specific polymerase chain reaction amplification is described using these primers and a conserved 3' primer corresponding to frame work 3, JH, or constant region. This strategy was used to isolate and sequence novel human germline VH genes belonging to the VH2 and VH4 families. Under certain conditions, chimeric VH sequences were created by a “jumping polymerase chain reaction”, combining DNA segments from different germline genes, but this could be avoided by limiting the number of amplification cycles. PCR amplification with these family specific primers will facilitate studies of the repertoire of germline VH genes as well as studies on VH gene usage in normal and aberrant (B cell malignancies, autoimmune diseases, etc.) B cell populations.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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