编码HLA - DR移植抗原α链的cDNA克隆的分离与鉴定。

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

作者信息

Gustafsson K, Bill P, Larhammar D, Wiman K, Claesson L, Schenning L, Servenius B, Sundelin J, Rask L, Peterson P A

出版信息

Scand J Immunol. 1982 Oct;16(4):303-8. doi: 10.1111/j.1365-3083.1982.tb00727.x.

DOI:10.1111/j.1365-3083.1982.tb00727.x

Abstract

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.

摘要

从拉吉细胞中分离出膜结合mRNA，并通过蔗糖梯度离心法富集编码HLA - DR移植抗原α链的信息。从该mRNA片段构建双链cDNA，连接到质粒pBR322上，并克隆到大肠杆菌中。通过杂交筛选，鉴定出一种能够与编码HLA - DRα链的mRNA杂交的质粒pDR - α - 1。从插入片段一端的核苷酸序列预测出的氨基酸序列，与从拉吉细胞中分离出的HLA - DRα链制剂的部分氨基末端序列相同。这清楚地表明pDR - α - 1携带了几乎完整的HLD - DRα链信息。从该质粒的核苷酸序列将有可能预测HLA - DRα链的一级结构。