title: [Cloning, expression and purification of human heat shock protein GP96 gene in Escherichia coli].
abstract: Heat shock protein gp96 is a member of HSP90 families. It can elicits both innate and adaptive immune responses. Generally it is essential to obtain enough amounts of pure gp96 to meet the needs of study and application. But the recombinant gp96 in E. coli is easy to degrade and form aggregates in certain conditions. In the experiment, first cloned human gp96 gene into pET-30a vector and expressed the recombinant in E. coli Blstar. Then purified gp96 by Ni-affinity column, anion exchange column and Gel-filtration in turn. In the end, removes most degraded fragments, aggregates and obtains certain amount of soluble gp96, which make an foundation for further investigations of the protein.
authors.affiliation: Institute of Microbiology Chinese Academy of Sciences, Beijing 100080, China. aliya-mongolia@yahoo.com.cn
authors.forename: A
authors.initials: A
authors.lastname: Li-Ya
authors.forename: Hong-Tao
authors.initials: HT
authors.lastname: Li
authors.forename: Ming-Hai
authors.initials: MH
authors.lastname: Zhou
authors.forename: Si
authors.initials: S
authors.lastname: Ha
authors.forename: Po
authors.initials: P
authors.lastname: Tien
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
chemical_list: D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; C053387:endoplasmin