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We have used a synthetic 20-nucleotide hybridization probe to isolate a cDNA clone encoding the alpha chain of the HLA-DR antigen from a cDNA library constructed from membrane-bound poly(A)+ mRNA. A set of synthetic 11-nucleotide fragments, potentially complementary to the codons for amino acids 11-14 of the HLA-DR alpha chain, were used to prime a cDNA synthesis reaction on various poly(A)+ mRNA templates. Extension of the primers in the presence of a single dideoxynucleotide triphosphate resulted in an 18-nucleotide cDNA product whose sequence corresponded to the NH2-terminal amino acids of the HLA-DR alpha chain. An oligonucleotide was synthesized based on this sequence information and its specificity for HLA-DR alpha mRNA was confirmed by primer extension and blot analysis. The cDNA library made from mRNA from the lymphoblastoid cell line CA-SC was probed with 32P-labeled cDNA synthesized on poly(A)+ mRNA from a B-cell line (CA-SC) or from a T-cell line (Molt-4) to enrich for B-cell-specific clones. A set of cDNA clones that hybridized preferentially with the B-cell probe was screened with the 32P-labeled 20-nucleotide probe. The cDNA clone isolated by this procedure is 1,100 nucleotides long; the nucleotide sequence of the 5' end of the cDNA insert corresponds to the amino acid sequence of the HLA-DR alpha chain. Hybridization of this cDNA clone to genomic blots suggests that the HLA-DR alpha chain is encoded by a single-copy gene. One of the restriction endonucleases used in genomic DNA digests reveals a restriction fragment polymorphism.
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
October 1982,
Scandinavian journal of immunology,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
January 1983,
DNA (Mary Ann Liebert, Inc.),
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
March 1982,
Proceedings of the National Academy of Sciences of the United States of America,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
February 1984,
Molecular biology & medicine,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
January 1982,
Proceedings of the National Academy of Sciences of the United States of America,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
September 1985,
Journal of immunology (Baltimore, Md. : 1950),
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
May 1985,
Annals of human genetics,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
January 1986,
Gene,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
October 1982,
Nature,
D Stetler, and
H Das, and
J H Nunberg, and
R Saiki, and
R Sheng-Dong, and
K B Mullis, and
S M Weissman, and
H A Erlich
November 1982,
Proceedings of the National Academy of Sciences of the United States of America,
