THE PRODUCTION OF RECOMBINANT HLA-DR-BETA AND INVARIANT CHAIN POLYPEPTIDES BY CDNA EXPRESSION IN ESCHERICHIA-COLI
THE PRODUCTION OF RECOMBINANT HLA-DR-BETA AND INVARIANT CHAIN POLYPEPTIDES BY CDNA EXPRESSION IN ESCHERICHIA-COLI
复制标题
DOI：
10.1016/0022-1759(87)90292-4
复制
发表时间：
1987-11-05
影响因子：
2.2
通讯作者：
KOCH, N
中科院分区：
文献类型：
--
作者：
KOCH, S;SCHULTZ, A;KOCH, N
在本报告中，我们描述了HLA-DRw 6 β的重组融合蛋白的生产。链和鼠Ia相关不变链。编码人HLA-DRw 6 β的cDNA。链和鼠Ia相关不变链被引入细菌表达质粒。这些质粒通过E.杆菌从粗E.大肠杆菌裂解物用于在兔中产生抗血清。这些抗血清能够免疫沉淀生物合成标记的II类和不变链抗原。另外，针对HLA-DR β的单个结构域产生了两种抗DR抗血清。链，从而产生具有确定的精细特异性的试剂。抗鼠不变链血清显示与人不变链交叉反应，因此可能有助于研究不同物种中不变链和Ia抗原的表达。这里描述的方法使我们能够生产大量的免疫相关蛋白质，用于生产多克隆和单克隆抗体。代表某些DR结构域的融合蛋白的可溶性片段也可用于功能免疫学研究。
In this report we describe the production of recombinant fusion proteins of the HLA-DRw6 .beta. chain and the murine Ia-associated invariant chain. cDNAs encoding the human HLA-DRw6 .beta. chain and the murine Ia-associated invariant chain were introduced into bacterial expression plasmids. These plasmids direct the synthesis of the respective molecules as fusion proteins of the bacteriophage MS-2 polymerase by E. coli. Fusion proteins purified from crude E. coli lysates were used to raise antisera in rabbits. These antisera were able to immunoprecipitate biosynthetically labelled class II and invariant chain antigens. Additionally, two anti-DR antisera were raised against single domains of the HLA-DR .beta. chain thus generating reagents with a defined fine specificity. The anti-murine invariant chain serum was shown to cross-react with the human invariant chain and therefore may be useful for studying invariant chain and Ia antigen expression in different species. The method described here permitted us to produce large quantities of immunologically relevant proteins, for use in the production of polyclonal and monoclonal antibodies. Soluble fragments of the fusion proteins representing certain DR domains may also be useful in functional immunological studies.