Construction of recombinant plasmid of Pseudomonas aeruginosa exotoxin A and its secreting expression in E. coli
- VernacularTitle:铜绿假单胞菌外毒素A表达载体的构建及可溶性表达
- Author:
Xiaomei HU
;
Xiancai RAO
;
Jianjun HUANG
- Publication Type:Journal Article
- Keywords:
Pseudomonas aeruginosa;
exotoxin A;
recombinant plasmid;
gene expression
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and to express the full-length Pseudomonas aeruginosa exotoxin A gene and to purify the expressed protein using affinity chromatography. Methods Exotoxin A gene was amplified from the recombinant plasmid pSK/PEA-T vector and subcloned into the pMAL-P2X vector. Then the recombinant plasmid pMAL- PEA was transformed to E.coli BL21. After induction with IPTG, the expressed protein was analyzed by SDS-PAGE and purified with affinity chromatography. Results The recombinants expressed the fusion protein at a level of about 20% of total cell proteins, and 80% of the fusion protein was secreted into the supernatant. Conclusion Successful expression and purification of PEA are of significance for in-depth study of the pathogenic mechanism of Pseudomonas aeruginosa and preparing immunotoxin.
