Isolation and identification of a cDNA clone coding for an HLA-DR transplantation antigen alpha-chain

• Published in: Scandinavian journal of immunology Vol. 16; no. 4; p. 303
• Main Authors: Gustafsson, K, Bill, P, Larhammar, D, Wiman, K, Claesson, L, Schenning, L, Servenius, B, Sundelin, J, Rask, L, Peterson, P A
• Format: Journal Article
• Language: English
• Published: England 01.10.1982
• Subjects: Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA - genetics; Genes; Genes, MHC Class II; HLA-DR Antigens; Humans; Plasmids
• ISSN: 0300-9475
• DOI: 10.1111/j.1365-3083.1982.tb00727.x

Membrane-bound mRNA was isolated from Raji cells and enriched for message coding for the HLA-DR transplantation antigen alpha-chain by sucrose gradient centrifugation. Double-stranded cDNA was constructed from this mRNA fraction, ligated to plasmid pBR322, and cloned into Escherichia coli. By hybrid selection, a plasmid, pDR-alpha-1, able to hybridize with mRNA coding for the HLA-DR alpha-chain was identified. From the nucleotide sequence of one end of the insert an amino acid sequence was predicted which is identical to part of the amino-terminal sequence of an HLA-DR alpha-chain preparation isolated from Raji cells. This clearly shows that pDR-alpha-1 carries almost the complete message for an HLD-DR alpha-chain. From the nucleotide sequence of this plasmid it will be possible to predict the primary structure of an HLA-DR alpha-chain.