人IgM Fab的噬菌体克隆及大肠杆菌表达
Bacteriophage cloning and Escherichia coli expression of a human IgM Fab.
作者信息
Hay B N, Sorge J A, Shopes B
机构信息
Stratacyte Corporation, La Jolla, CA 92037.
出版信息
Hum Antibodies Hybridomas. 1992 Apr;3(2):81-5.
PMID:1385989
Abstract
We have combined the molecular biology methods of the polymerase chain reaction and recombinant DNA cloning in bacteriophage lambda to express a human IgM Fab in Escherichia coli using genes derived from an Epstein-Barr virus transformed cell line. This method comprises three cDNA amplifications and a single cloning step, culminating in the stable overexpression of mammalian heterodimeric recombinant protein in a prokaryotic host.
摘要
我们将聚合酶链反应的分子生物学方法与噬菌体λ中的重组DNA克隆技术相结合，利用源自爱泼斯坦-巴尔病毒转化细胞系的基因在大肠杆菌中表达人IgM Fab。该方法包括三次cDNA扩增和一个单一的克隆步骤，最终在原核宿主中稳定过量表达哺乳动物异二聚体重组蛋白。
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