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一种EF-1α样大鼠cDNA(S1)在大肠杆菌中的表达以及针对重组蛋白的兔多克隆抗血清的制备。

Expression of an EF-1 alpha-like rat cDNA, S1, in E. coli and production of a rabbit polyclonal antiserum to the recombinant protein.

作者信息

Liu C H, Liu S, Wang E

机构信息

Bloomfield Centre for Research in Aging, Lady Davis Institute for Medical Research, Jewish General Hospital, Montréal, Québec, Canada.

出版信息

Biochem Biophys Res Commun. 1993 Sep 30;195(3):1371-8. doi: 10.1006/bbrc.1993.2195.

Abstract

A previously identified rat cDNA (S1) that shares 78% nucleotide homology and a predicted 92% amino acid homology with human EF-1 alpha was constructed into PGEX-2T and inducibly expressed in E. coli as a fusion protein. The purified recombinant S1 protein (rpS1) was used to raise rabbit antiserum which recognized rpS1 but not EF-1 alpha on immunoblots. A polyclonal antiserum to EF-1 alpha failed to react with the rpS1. Our results indicate that rpS1 protein can be used to produce antibodies which distinguish pS1 from EF-1 alpha despite their extensive amino acid sequence homology.

摘要

一个先前鉴定出的大鼠cDNA(S1),它与人类EF-1α具有78%的核苷酸同源性以及预测的92%的氨基酸同源性,被构建到PGEX-2T载体中,并在大肠杆菌中诱导表达为融合蛋白。纯化的重组S1蛋白(rpS1)被用于制备兔抗血清,该抗血清在免疫印迹中能识别rpS1而不识别EF-1α。针对EF-1α的多克隆抗血清与rpS1不发生反应。我们的结果表明,尽管rpS1蛋白与EF-1α具有广泛的氨基酸序列同源性,但它可用于产生能区分rpS1和EF-1α的抗体。

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