Soluble expression of recombinant human apoliprotein A-I-Milano in Escherichia coli.
- Author:
Ming LI
1
;
Hong-Liang ZHAO
;
Chong XUE
;
Wei ZHANG
;
Shi-Meng ZHANG
;
Zhi-Min LIU
Author Information
1. Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China. liming096@sina.com
- Publication Type:Journal Article
- MeSH:
Apolipoprotein A-I;
biosynthesis;
genetics;
Escherichia coli;
genetics;
metabolism;
Humans;
Mutant Proteins;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
Solubility
- From:Chinese Journal of Biotechnology
2005;21(3):354-359
- CountryChina
- Language:Chinese
-
Abstract:
Apolipoprotein A-I-Milano(AIM), a natural variant, not only inhibits the initiation and progression of atherosclerosis, but also makes the preexisting atherosclerotic lesions regress. AIM gene, at which N-terminal codens were optimized, was subcloned into the expression vector of pET22b. Recombiant plasmids were transformed into E. coli strain BL21 (DE3) and induced with IPTG. The expressed apoliprotein A-I-Milano was soluble in E. coli and was about 38% of total cell lysate. Purified by Butyl Sepharose 4F. F hydrophobic chromatography and Q Sepharose H.P. anion exchange chromatography, followed by ultrafiltration with Vivaspin 20 (30 000MW), AIM monomer was obtained in a purity of more than 95%. Activity assay of binding of AIM monomer to lipid indicates that association of AIM monomer with DMPC is slower than normal apoA-I but DMPC number associated by AIM monomer is more than by apoA-I. This results will be important for studying structure, function of AIM, specially clinical application.
