title: [High level expression and purification of Cysticercosis cellulose annexin32 in Escherichia coli].
abstract: In previous work, the cDNA encoding Cysticercus cellulose annexin32 has been cloned. With PCR method, two different restriction Sites were added to each end of the cDNA respectively. Then, the cDNA was inserted into prokaryotic expression vector pJLA-503. After inducing, most foreign protein was expressed in soluble form, which was up to 35% of the total protein of the bacteria. Subsequently, the recombinant Annexin32 was purified with (NH4)2SO4 stepwise precipitation, DEAE-Sepharose FF and Sephacryl S-200 HR chromatography. The final pure protein can been shown as a single band in SDS-PAGE, and the biological activity was verified by Western blot and anticoagulation activity assay.
authors.affiliation: Department of Medical Genetics, Second Military Medical University, Shanghai 200433, China. guo-yingjun@263.net
authors.forename: Y J
authors.initials: YJ
authors.lastname: Guo
authors.forename: D
authors.initials: D
authors.lastname: Wu
authors.forename: H L
authors.initials: HL
authors.lastname: Yan
authors.forename: S H
authors.initials: SH
authors.lastname: Sun
mesh_terms: D000818:Animals; D017302:Annexins / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D000925:Anticoagulants / Q000494:pharmacology; D015153:Blotting, Western; D003552:Cysticercus / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D015801:Helminth Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D017302:Annexins; D000925:Anticoagulants; D018076:DNA, Complementary; D015801:Helminth Proteins; D011994:Recombinant Proteins