Isolation and Identification of a cDNA Clone Coding for an HLA-DR Transplantation Antigen alpha-Chain

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Abstract
Membrane‐bound mRNA was isolated from Raji cells and enriched for message coding for the HLA‐DR transplantation antigen α‐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‐α‐1, able to hybridize with mRNA coding for the HLA‐DR α‐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 α‐chain preparation isolated from Raji cells. This clearly shows that pDR‐α‐1 carries almost the complete message for an HLA‐DR α‐cbain. From the nucleotide sequence of this plasmid it will be possible to predict the primary structure of an HLA‐DR α‐chain.