PMID: 6403940Mar 1, 1983Paper

Use of synthetic oligonucleotide probes complementary to genes for human HLA-DR alpha and beta as extension primers for the isolation of 5'-specific genomic clones

Proceedings of the National Academy of Sciences of the United States of America
H K DasS M Weissman

Abstract

We have synthesized 175-nucleotide-long probes for the DNA of human histocompatibility antigens HLA-DR alpha and beta by extending on poly(A)+ mRNA from B-cell lines with short synthetic deoxyribonucleotide primers complementary to the predicted nucleotide sequence of the NH2 terminus of both polypeptides. The synthesis of the probe for the alpha-chain DNA was a two-step process starting with 11-mers which were extended by dideoxynucleotide chain termination experiments to a 20-mer of predicted sequence. The synthesized 20-mer was then used to generate a 175-nucleotide cDNA probe which was shown to encode the appropriate amino acids for the alpha chain and was used to select a human genomic DNA clone containing the coding sequences for HLA-DR alpha. For the beta polypeptide an 18-mer homologous to the NH2-terminal sequence of a cDNA clone from another B-cell line was used to extend on poly(A)+ mRNA isolated from a B-cell line. Preliminary sequence analysis of a 175-base-long extension product indicates a match of the cDNA sequence to the published sequence of a clone for HLA-DR beta. Information from these extension experiments helps to establish the sensitivity and specificity of the primer extension method.

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Citations

Jul 15, 1999·Computers & Chemistry·Y Ding, C E Lawrence
Jun 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·H K DasS M Weissman
May 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·J I BellH O McDevitt
Dec 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·U A Germann, K Lerch
Jul 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·M A BlanarR A Flavell
Oct 25, 1985·Nucleic Acids Research·S K LawranceS M Weissman
Jan 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·N K ZellerR A Lazzarini
Apr 1, 1986·Medicinal Research Reviews·S P Adams, G R Galluppi
Jan 1, 1987·Annals of the New York Academy of Sciences·G A Vehar

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