Cloning the heavy chain of human HLA-DR antigen using synthetic oligodeoxyribonucleotides as hybridization probes

• Published in: DNA (New York, N.Y.) Vol. 2; no. 3; p. 175
• Main Authors: Kajimura, Y, Toyoda, H, Sato, M, Miyakoshi, S, Kaplan, S A, Ike, Y, Goyert, S M, Silver, J, Hawke, D, Shively, J E
• Format: Journal Article
• Language: English
• Published: United States 01-01-1983
• Subjects: Amino Acid Sequence; Base Sequence; Cell Line; Cloning, Molecular; Genes, MHC Class II; Genes, Synthetic; HLA-DR Antigens; Humans; Nucleic Acid Hybridization; Oligodeoxyribonucleotides - genetics; RNA, Messenger - genetics
• ISSN: 0198-0238
• DOI: 10.1089/dna.1983.2.175

The recent development of the amino acid microsequence technique allows us to obtain partial sequence information using an extremely small amount of protein. Two sets of mixed oligonucleotide probes were chemically synthesized using the amino acid sequence information for the heavy chain of human HLA-DR antigen obtained by the microsequence technique. These two hybridization probes were used to screen cDNA clones constructed from cytoplasmic poly(A)+ mRNA from a human B lymphoblastoid homozygous cell line (LG-2). Of the 10,000 clones screened, two clones hybridized with the probes. DNA sequence analysis showed that the longer one of the two cDNA clones was 1183 nucleotides long, including the entire coding region, the signal peptide region, and the complete 3'-noncoding region. The deduced amino acid sequence of the HLA-DR alpha chain is identical to that of other cell lines with a different HLA-DR typing. However, several nucleotide differences are found in the 3'-untranslated region compared with that of other DR haplotypes.