Optimization of expression of an annexin V-hirudin chimeric protein in Escherichia coli.
A human Annexin V-Hirudin chimeric protein, Annexin V-Hirudin C, was expressed in Escherichia coli. A broad range of parameters such as plasmid stability during propogation and expression, expression capacity stability, the culture media, the growth time before induction and the induction duration were examined and optimized. Recombinant Annexin V-Hirudin C was purified from the cell lysate supernatants by ethanol precipitation, DEAE-cellulose chromatography and Sephadex G-75 chromatography, and the purified protein showed dose-dependent thrombin inhibitory activity. The overall production of purified Annexin V-Hirudin C protein is 10 mg/l/OD600.
State Key Laboratory of Pharmaceutical Biotechnology, Institute of Molecular and Cell Biology, College of Life Sciences, Nanjing University, Nanjing 210093, PR China.
Hui
H
Yuan
Xiang
X
Yang
Zi-Chun
ZC
Hua
D017304:Annexin A5 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001709:Biotechnology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006629:Hirudins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D017304:Annexin A5; D006629:Hirudins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins