title: [Soluble expression of recombinant human apoliprotein A-I-Milano in Escherichia coli].
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.
authors.affiliation: Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China. liming096@sina.com
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.forename: Hong-Liang
authors.initials: HL
authors.lastname: Zhao
authors.forename: Chong
authors.initials: C
authors.lastname: Xue
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Shi-Meng
authors.initials: SM
authors.lastname: Zhang
authors.forename: Zhi-Min
authors.initials: ZM
authors.lastname: Liu
mesh_terms: D016632:Apolipoprotein A-I / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D050505:Mutant Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012995:Solubility
chemical_list: D016632:Apolipoprotein A-I; D050505:Mutant Proteins; D011994:Recombinant Proteins