AN EXPRESSION SYSTEM FOR SECRETION AND PURIFICATION OF A GENETICALLY-ENGINEERED THERMOSTABLE CHIMERA OF PROTEIN-A AND ALKALINE-PHOSPHATASE

被引：11

作者：

CHOWDHURY, PS ^{[1
]}

KUSHWAHA, A ^{[1
]}

ABROL, S ^{[1
]}

CHAUDHARY, VK ^{[1
]}

机构：

[1] UNIV DELHI,DEPT BIOCHEM,NEW DELHI 110021,INDIA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1994年 / 5卷 / 01期

关键词：

D O I：

10.1006/prep.1994.1013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A chimera between gene segments of Protein A and a mutated alkaline phosphatase (lysine328 mutated to alanine) of Escherichia coli has been constructed. This chimeric gene was cloned in a T7 promoter-based IPTG-inducible expression vector. The chimeric protein was expressed in E. coli and was efficiently secreted into the periplasm, from which it could he easily purified by a combination of ion-exchange and gel permeation chromatography. The purified chimera was found to be thermostable and exhibited both IgG binding and high alkaline phosphatase activity. It was used as a probe in enzyme-linked immunosorbent assays and results indicate that it is a promising substitute for secondary antibodies in enzyme-linked immunoassays. © 1994 Academic Press. All rights reserved.

引用

页码：89 / 95

页数：7

共 28 条

[11] PROTEIN A FROM STAPHYLOCOCCUS-AUREUS - ITS ISOLATION BY AFFINITY CHROMATOGRAPHY AND ITS USE AS AN IMMUNOSORBENT FOR ISOLATION OF IMMUNOGLOBULINS [J].