Soluble expression of recombinant human apoliprotein A-I-Milano in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 354-359, 2005.
Article
in Zh
| WPRIM
| ID: wpr-305270
Responsible library:
WPRO
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.
Full text:
1
Database:
WPRIM
Main subject:
Solubility
/
Recombinant Proteins
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Apolipoprotein A-I
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Escherichia coli
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Mutant Proteins
/
Genetics
/
Metabolism
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Biotechnology
Year:
2005
Document type:
Article