2006•Zhongguo shouyi xuebaoRequires access
cDNA Cloning of FMDV Structural Protein VP2-3-1 Gene and its Prokaryotic Expression
Zaixin Liu
Open publisher page 2 citations
Abstract
FMDV P1 gene was amplified from positive plasmid pGEM-p1. PCR product of P1 was digested with NcoⅠ and XhoⅠ, and got objective gene VP2-3-1. This gene was ligated into vector pProEX-HTb and digested with NcoⅠ and XhoⅠ respectively, then transformed into E.coli BL21(DE3) cells. The recombinant plasmid were identified by restriction analysis and PCR and DNA sequencing. A recombinant plasmid with FMDV VP2-3-1 gene was selected and named as pProEX-VP2-3-1. The bacteria containing pProEX-VP2-3-1 was induced with Isopropyl-D-galactoside(IPTG) and the bacteria culture fluid were sampled and examined by SDS-PAGE and Western blotting after being properly treated. The results showed that the VP2-3-1 gene was successfully expressed in E.coli and could be recognized by the positive bovine serum to FMDV.