Construction of indirect ELISA method using rabies virus G gene expressed in prokaryotic system
LISheng-li
YUXiang-dong
WANGWei-jie
HUYi-yuan
Abstract：In order to acquire rabies virus G gene and express it in E.coli and establish the evaluation method for rabies virus immune,according to Rabies virus ERA G gene sequence to design primer se-quences,RNA was extracted from rabies virus,the first strand cDNA was synthesized by reverse transcrip-tion.G gene fragments were amplified by PCR.Then the gene fragment was cloned to the prokaryotic ex-pression vector pET28a (+)to construct PET-RVG.The positive recombinant plasmids were transfected into E.coli BL21(DE3)gold,with IPTG induced expression and determined the best expression condition. Using Niaffinity chromatography purification with G protein,and the indirect ELISA assay for the detection of rabies virus antibodies in serum was established based on the G protein.The results showed that the puri-ty degree was 96%.
Keywords：rabies virusG geneprokaryotic expressionindirect enzyme-linked immunosorbent assay (ELISA)evaluation of immune
Publication Date：2014-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 24-28 )