Isolation of the Gene Encoding the Human T-lymphocyte Differentiation Antigen Leu-2 (T8) by Gene Transfer and cDNA Subtraction
We report the isolation of genomic and cDNA clones encoding the human T-lymphocyte cell-surface differentiation antigen, Leu-2 (T8), by use of a combination of transfection, fluorescence-activated cell-sorting, and subtractive cDNA hybridization. We constructed a cDNA library with mRNA from a mouse...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1984-12, Vol.81 (24), p.7688-7692 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Kavathas, Paula Sukhatme, Vikas P. Herzenberg, Leonard A. Parnes, Jane R. |
| description | We report the isolation of genomic and cDNA clones encoding the human T-lymphocyte cell-surface differentiation antigen, Leu-2 (T8), by use of a combination of transfection, fluorescence-activated cell-sorting, and subtractive cDNA hybridization. We constructed a cDNA library with mRNA from a mouse L-cell transfectant in which the human Leu-2 gene is expressed and amplified. We identified Leu-2 cDNA clones by screening with a selected cDNA probe from a second amplified Leu-2 transfectant. This probe contained cDNA species not removed by hybridization with L-cell mRNA. A Leu-2 cDNA clone was used to isolate a genomic clone. Transfection with DNA from this clone resulted in a high number of Leu-2 transfectants. This approach can be used to clone genes coding for other cell-surface molecules. |
| doi_str_mv | 10.1073/pnas.81.24.7688 |
| format | Article |
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We constructed a cDNA library with mRNA from a mouse L-cell transfectant in which the human Leu-2 gene is expressed and amplified. We identified Leu-2 cDNA clones by screening with a selected cDNA probe from a second amplified Leu-2 transfectant. This probe contained cDNA species not removed by hybridization with L-cell mRNA. A Leu-2 cDNA clone was used to isolate a genomic clone. Transfection with DNA from this clone resulted in a high number of Leu-2 transfectants. 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We constructed a cDNA library with mRNA from a mouse L-cell transfectant in which the human Leu-2 gene is expressed and amplified. We identified Leu-2 cDNA clones by screening with a selected cDNA probe from a second amplified Leu-2 transfectant. This probe contained cDNA species not removed by hybridization with L-cell mRNA. A Leu-2 cDNA clone was used to isolate a genomic clone. Transfection with DNA from this clone resulted in a high number of Leu-2 transfectants. This approach can be used to clone genes coding for other cell-surface molecules.</description><subject>Animals</subject><subject>Antigens, Differentiation, T-Lymphocyte</subject><subject>Antigens, Surface - genetics</subject><subject>CDNA libraries</subject><subject>Cell lines</subject><subject>Coliphages - genetics</subject><subject>Complementary DNA</subject><subject>DNA</subject><subject>DNA - metabolism</subject><subject>Escherichia coli - genetics</subject><subject>Gene Amplification</subject><subject>Genes</subject><subject>Genetic hybridization</subject><subject>Genomics</subject><subject>Humans</subject><subject>L cells</subject><subject>L Cells (Cell Line)</subject><subject>Messenger RNA</subject><subject>Mice</subject><subject>Nucleic Acid Hybridization</subject><subject>Plasmids</subject><subject>RNA</subject><subject>RNA, Messenger - genetics</subject><subject>T lymphocytes</subject><subject>T-Lymphocytes - immunology</subject><subject>Transfection</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc2P0zAUxC0EWsrCGQkJ5BPsHtK1HTtxDhyq_ZYqOFDOlus8t1kldrETRC_87TiklOXCyZbnN_OeNQi9pmROSZlf7JyOc0nnjM_LQsonaEZJRbOCV-QpmhHCykxyxp-jFzE-EEIqIckJOik4J5SzGfp5H32r-8Y77C3ut4BvwQG-dsbXjdv8frkbOu3wKmv33W7rzb4HfNVYCwFc30zeRbptwOElDBnDZyt5jtf7KWoVtIsJxtrV2Fx9WuAvw7oP2ozGl-iZ1W2EV4fzFH29uV5d3mXLz7f3l4tlZvL0rSwXTFMrrCWSlEBobQuqNRBRQMHqiteWVVByKkRBQHBTGmsrziXUYKyhLD9FH6fc3bDuoDZp86BbtQtNp8Need2ofxXXbNXGf1d5xRgtk__9wR_8twFir7omGmhb7cAPUZVCskIWNIEXE2iCjzGAPc6gRI2NqbExJaliXI2NJcfbx6sd-UNFSf9w0EfjH_VvgLJD2_bwo0_ku_-SCXgzAQ-x9-FIsDRJ5L8Ao860RA</recordid><startdate>19841201</startdate><enddate>19841201</enddate><creator>Kavathas, Paula</creator><creator>Sukhatme, Vikas P.</creator><creator>Herzenberg, Leonard A.</creator><creator>Parnes, Jane R.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19841201</creationdate><title>Isolation of the Gene Encoding the Human T-lymphocyte Differentiation Antigen Leu-2 (T8) by Gene Transfer and cDNA Subtraction</title><author>Kavathas, Paula ; Sukhatme, Vikas P. ; Herzenberg, Leonard A. ; Parnes, Jane R.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3768-352a1f5ff0807e01df61aae056e62d94df29e7415560e54c7cff9448edecfc123</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Animals</topic><topic>Antigens, Differentiation, T-Lymphocyte</topic><topic>Antigens, Surface - genetics</topic><topic>CDNA libraries</topic><topic>Cell lines</topic><topic>Coliphages - genetics</topic><topic>Complementary DNA</topic><topic>DNA</topic><topic>DNA - metabolism</topic><topic>Escherichia coli - genetics</topic><topic>Gene Amplification</topic><topic>Genes</topic><topic>Genetic hybridization</topic><topic>Genomics</topic><topic>Humans</topic><topic>L cells</topic><topic>L Cells (Cell Line)</topic><topic>Messenger RNA</topic><topic>Mice</topic><topic>Nucleic Acid Hybridization</topic><topic>Plasmids</topic><topic>RNA</topic><topic>RNA, Messenger - genetics</topic><topic>T lymphocytes</topic><topic>T-Lymphocytes - immunology</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kavathas, Paula</creatorcontrib><creatorcontrib>Sukhatme, Vikas P.</creatorcontrib><creatorcontrib>Herzenberg, Leonard A.</creatorcontrib><creatorcontrib>Parnes, Jane R.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kavathas, Paula</au><au>Sukhatme, Vikas P.</au><au>Herzenberg, Leonard A.</au><au>Parnes, Jane R.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation of the Gene Encoding the Human T-lymphocyte Differentiation Antigen Leu-2 (T8) by Gene Transfer and cDNA Subtraction</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1984-12-01</date><risdate>1984</risdate><volume>81</volume><issue>24</issue><spage>7688</spage><epage>7692</epage><pages>7688-7692</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>We report the isolation of genomic and cDNA clones encoding the human T-lymphocyte cell-surface differentiation antigen, Leu-2 (T8), by use of a combination of transfection, fluorescence-activated cell-sorting, and subtractive cDNA hybridization. We constructed a cDNA library with mRNA from a mouse L-cell transfectant in which the human Leu-2 gene is expressed and amplified. We identified Leu-2 cDNA clones by screening with a selected cDNA probe from a second amplified Leu-2 transfectant. This probe contained cDNA species not removed by hybridization with L-cell mRNA. A Leu-2 cDNA clone was used to isolate a genomic clone. Transfection with DNA from this clone resulted in a high number of Leu-2 transfectants. This approach can be used to clone genes coding for other cell-surface molecules.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>6440142</pmid><doi>10.1073/pnas.81.24.7688</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0027-8424 |
| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1984-12, Vol.81 (24), p.7688-7692 |
| issn | 0027-8424 1091-6490 |
| language | eng |
| recordid | cdi_pubmed_primary_6440142 |
| source | MEDLINE; JSTOR Archive Collection A-Z Listing; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | Animals Antigens, Differentiation, T-Lymphocyte Antigens, Surface - genetics CDNA libraries Cell lines Coliphages - genetics Complementary DNA DNA DNA - metabolism Escherichia coli - genetics Gene Amplification Genes Genetic hybridization Genomics Humans L cells L Cells (Cell Line) Messenger RNA Mice Nucleic Acid Hybridization Plasmids RNA RNA, Messenger - genetics T lymphocytes T-Lymphocytes - immunology Transfection |
| title | Isolation of the Gene Encoding the Human T-lymphocyte Differentiation Antigen Leu-2 (T8) by Gene Transfer and cDNA Subtraction |
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