Chinese Journal of Veterinary Science 2009;29(7):836-840

Cloning and expression of the σC gene of avian reovirus

Xiaobo HUANG ; Shanshan LING ; Sanjie CAO ; Xintian WEN ; Cunwei WANG ; Yuankun CHEN

Keywords

avian reovirus; σC gene; pMD19T-σC; pET32a-σC

Country

China

Language

Chinese

Abstract

The σC gene of ARV S1133 was designed to amplify by reverse transcription chain reaction(RTPCR).The σC gene was inserted into the vector pMD19-T,identified by PCR method and restriction enzyme,and sequenced.It showed that the insert cloned gene fragment was the σC gene of ARV.Then the gene was inserted intc the pET32a(+) and indicated that fusion expression vector pET32a-σC was constructed.The recombinant fusion protein was highly expressed in E.coli BL21 induced by 1.0 mmol/L IPTG for 5 hours in the form of inclusion bodies.The weight of recombinant fusion protein molecular is 54 000.Western-blot with ARV antibodies against the fusion protein showed the recombinant protein has a favourable reactivity.