Preparation and physiological functions identification of JC virus VLP-Z
10.3760/cma.j.issn.0254-5101.2009.12.017
- VernacularTitle:JC病毒VLP-Z的制备及其生物活性鉴定
- Author:
Haiyan ZHAI
;
Qiumin QU
;
Xiao ZHE
- Publication Type:Journal Article
- Keywords:
JC virus;
Virus-like particle;
Gene therapy;
Target gene delivery
- From:Chinese Journal of Microbiology and Immunology
2009;29(12):1114-1117
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express the recombinant protein VP1-Z, and investigate whether VLP-Z has the physiological functions like as wild-type VLP. Methods The expression plasmid pET15b-VP1-Z was introduced into competent E. coil BL21 (DF3)/pLys cells by transformation, and the expression of re-combinant protein VP1-Z was induced by incubation of the cells with IPTG. The protein was prepared as pre-viously described for wild-type VLP. The morphous of VLP-Z were observed by electron microscopy, and the physiological functions of VLP-Z were investigated by hemagglutination test and by immunofluorescence. Re-sults The purified VLP-Z composed of VP1-Z possessed hemagglutination activity and yielded a prominent band of 50×10~3 on SDS-PAGE and staining with Coomassie Brilliant Blue. The VLP-Z exhibited virus-like particles structure like as wild-type VLP with a diameter of 45-50 nm, which was slightly bigger than that of wild-type VLP(42-45 nm). In immunofluorescence test, VP1-Z was detected within the cytoplasm and nu-cleus after HeLa cells were inoculated with VLP-Z. Conclusion The physiological functions of recombinat-ed protein VLP-Z were comparable with wild-type VLP.
