Expression and purification of recombinant JEV EDⅢ protein and its application in Array-ELISA assay
10.3760/cma.j.issn.0254-5101.2013.12.013
- VernacularTitle:乙型脑炎病毒 EDⅢ蛋白的表达纯化及在 Array-ELISA 方法中的应用
- Author:
Yang ZHENG
;
Xiaoping KANG
;
Yuchang LI
;
Xiaoyan WU
;
Xiaosong ZHANG
;
Yinhui YANG
- Publication Type:Journal Article
- Keywords:
Japanese encephalitis virus;
EDⅢprotein;
Clone;
Array-ELISA
- From:Chinese Journal of Microbiology and Immunology
2013;(12):954-959
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express and purify Japanese encephalitis virus ( JEV) EDⅢprotein and evaluate the possibility of using it as a candidate antigen in JEV diagnostic kit .Methods PCR primers spe-cific for the gene encoding JEV EDⅢprotein were designed and used to amplify the gene fragment by RT-PCR.The cloned gene fragment was then inserted into pET-30a (+) to construct the recombinant expression plasmid.The transformed E.coli BL21 carrying expression plasmid were induced by IPTG to express JEV EDⅢ protein.The expressed JEV EDⅢprotein and a control antigen of tick-borne encephalitis virus protein were deposited in small spots to set up ELISA microarray .The serum samples from patients with Japanese encephalitis and healthy people were detected by Array-ELISA.The results obtained by Array-ELISA were compared with those by using indirect immunofluorescence assay .Results The gene fragment encoding JEV EDⅢprotein was successfully cloned and expressed in E.coli BL21.The recombinant protein could be used in Array-ELISA assay for the detection of serum samples from patients with Japanese encephalitis and healthy subjects .The results were consistent with those by using indirect immunofluorescence assay.Conclusion The recombinant JEV EDⅢprotein can be used as a candidate antigen for the diagnosis of JEV infection .
