Expression of t antigen fusion protein of JC virus and preparation of its polyclonal antibody
10.3760/cma.j.issn.1000-6680.2009.07.005
- VernacularTitle:JC病毒t抗原的融合蛋白表达及抗体制备
- Author:
Tielong ZHENG
;
Dianli WANG
;
Xingwang LI
;
Yu MAO
;
Yuan HONG
;
Qi WANG
;
Jun CHENG
- Publication Type:Journal Article
- Keywords:
JC virus;
Antigens,viral,tumor;
Antibody formation;
Gene expression;
Recombinant fusion proteins
- From:
Chinese Journal of Infectious Diseases
2009;27(7):403-407
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct prokaryotic expression vector carrying jc virus(JCV)t-antigen gene,express and purify this fusion protein.Methods The JCV t-antigen gene from a cerebrospinal fluid sample was amplified using polymerase chain reaction(PCR)method.After sequencing.the gene was cloned into plasmid pET32a(+)to construct recombinant prokaryotic expression vector pET32a(+)-t.The t-antigen fusion protein was expressed by isopropy-~D-thiogalactoside(IPTG)induction and prepared in large scale,then purified by Ni+affinity column chromatography.The polyclonal antibody was obtained from the BAI.B/C mouse immunity by the purified protein.Results The relative molecular nlass of recombinant protein expressed by pET32a(+)-t was about 41 000.Sodium dodeeylsulfate-polyaerylamide gel electrophoresis(SDS-PAGE)showed that the fusion protein W&S highly expressed after 3.5～20.Oh of IPTG induction.The antigenicity of the purified protein Was well confirmed by Western blot.The anti-mousepolyclonal antibody was obtained successfully from immunized BALB/c mice.Conclusions The prokaryotic expression vector pET32a(+)-t is successful constructed and the fusion protein is expressed and purified.Furthermore,the antibody of JCV small envelop protein t is successfully prepared.This work provides vMuable information for further study on epidemiology and biological function of t antigen.
