EXPRESSION OF TICK-BORNE ENCEPHALITIS VIRUS PRM-E PROTEIN IN EUKARYOTIC SYSTEM AND ITS APPLICATION IN LUCIFERASE IMMUNOPRECIPITATION SYSTEM
10.3969/j.issn.1005-0507.2016.01.007
- VernacularTitle:蜱传脑炎病毒prM-E蛋白的真核表达及其在荧光素酶免疫共沉淀技术中的应用?
- Author:
Xin RAN
1
;
XiaoPing KANG
;
YuChang LI
;
XiaoYan WU
;
NaiFan HUO
;
Jia JIA
;
XueFeng CAO
;
YinHui YANG
Author Information
1. 安徽医科大学
- Keywords:
Tick-borne encephalitis virus;
prM-E protein;
Eukaryotic expression;
Diagnostic antigen
- From:
Acta Parasitologica et Medica Entomologica Sinica
2016;23(1):39-44
- CountryChina
- Language:Chinese
-
Abstract:
In order to establish a novel serologic test—Luciferase immunoprecipitation system ( LIPS) , full?length prM?E of TBE virus was expressed by eukaryotic expression system and its effect was evaluated by LIPS technology. Firstly, using TBEV′s RNA as template, prM?E gene of TBEV was amplified by RT?PCR, eukaryotic expression vector pNLF?prM?E was constructed through inserting prM?E gene into pNLF1?secN vector. Fusion protein prM?E?luciferase was expressed by transfecting Cos7 cells. The expression of fusion protein was measured by testing values of luciferase in supernatant of Cos7 cells. Immunofluorescence assay ( IFA) was employed to identify the specificity of the recombinant protein and LIPS was used to detect the serum of TBEV infected patients. High expression level of luciferase was tested in the supernatants of the transfected cells. The specific binding of prM?E with TBEV antibody was confirmed by IFA. The LIPS assay detected 19 positive results out of 20 sera of the TBEV infected patients, 7 control sera all showed negative results. The results indicated that the expressed prM?E had good sensitivity and specitivity, which could be used as a candidate diagnostic antigen for LIPS assay for TBEV infection.
