Cloning of UL7 protein of HSV-1 and analysis of the expression of UL7 protein during the prolifera-tion of HSV-1
10.3760/cma.j.issn.0254-5101.2016.04.001
- VernacularTitle:HSV-1衣被蛋白UL7基因的克隆及其在病毒增殖中的表达分析
- Author:
Jienan ZHOU
;
Xingli XU
;
You GAO
;
Ying ZHANG
;
Jingjing WANG
;
Longding LIU
;
Qihan LI
- Publication Type:Journal Article
- Keywords:
Herpes simplex virus 1 (HSV-1);
UL7;
Prokaryotic expression;
Polyclonal antibody
- From:
Chinese Journal of Microbiology and Immunology
2016;36(4):241-246
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express and purify the recombinant UL7 protein of herpes simplex virus 1 (HSV-1), to prepare the corresponding UL7-specific polyclonal antibody and to preliminarily analyze the expression of UL7 protein during the proliferation of HSV-1. Methods The UL7 gene was amplified by PCR and then cloned into the pGEX-5X-1 vector for expression of UL7 protein in the prokaryotic expression system. The constructed expression plasmid, pGEX-5X-1-UL7, was transformed into E. coli BL21 (DE3) to induce the expression of UL7 protein by IPTG. The purified GST-UL7 fusion protein was used as antigen to inject the ICR mouse for the preparation of polyclonal antibody specific for UL7 protein. The titer and speci-ficity of the polyclonal antibody were analyzed by using indirect ELISA and Western blot assay, respectively. The UL7 protein-specific polyclonal antibody was used to detect the expression of UL7 protein at different time points after infecting Vero cells with HSV-1. Results The GST-UL7 fusion protein was efficiently ex-pressed in E. coli BL21 (DE3). The UL7 protein-specific polyclonal antibody was prepared with high titer (1 ∶ 105) and high specificity as indicated by the indirect ELISA and Western blot assay. The expression of UL7 protein was detected at different time points after infecting Vero cells with HSV-1. Conclusion The GST-UL7 fusion protein was obtained successfully and the UL7 protein-specific polyclonal antibody was pre-pared. Accompany with the proliferation of HSV-1, the expression of UL7 protein was detected at different time points by using the polyclonal antibody.
