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Isolation of Distinct cDNA Clones Encoding HLA-DR β Chains by Use of an Expression Assay
Eric O. Long, Claire T. Wake, Michel Strubin, Nicole Gross, Roberto S. Accolla, Stefan Carrel and Bernard Mach
Proceedings of the National Academy of Sciences of the United States of America
Vol. 79, No. 23, [Part 1: Biological Sciences] (Dec. 1, 1982), pp. 7465-7469
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/13108
Page Count: 5
You can always find the topics here!Topics: Complementary DNA, Messenger RNA, Cell lines, B lymphocytes, Oocytes, RNA, Antigens, Monoclonal antibodies, T lymphocytes, Amino acids
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cDNA clones encoding different human Ia antigen β chains were isolated by use of a complementation-expression assay in Xenopus oocytes. The assay was based on two previous findings. First, oocytes injected with mRNA from a human B-cell line express HLA-DR antigen. The three intracellular DR chains are assembled in oocytes and can be immunoprecipitated with anti-DR monoclonal antibodies. Second, we have isolated cDNA clones encoding DR α and intermediate chains. In order to identify β -chain cDNA clones, mRNA was hybrid-selected with pools of cDNA clones, mixed with mRNA for the α and intermediate chains, and injected into oocytes. We isolated two distinct clones that could select DR β -chain mRNA as demonstrated by assembly of the translation product with DR α chains and immunoprecipitation with DR-specific monoclonal antibodies. One clone is specific for a β chain of the DR locus. The other clone, much weaker in its ability to select DR mRNA, encodes another Ia-like β chain. Full-length cDNA clones corresponding to the DR and Ia-like β chains were isolated and compared. Cross-hybridization was detectable in the coding regions but not in the 3′ untranslated regions. Distinct RNAs homologous to the DR and the Ia-like β -chain clones were present in B cells but were undetectable in three T-cell lines.
Proceedings of the National Academy of Sciences of the United States of America © 1982 National Academy of Sciences