Cloning and expression of the σC gene of avian reovirus
- VernacularTitle:禽呼肠孤病毒σC基因的克隆和表达
- Author:
Xiaobo HUANG
;
Shanshan LING
;
Sanjie CAO
;
Xintian WEN
;
Cunwei WANG
;
Yuankun CHEN
- Publication Type:Journal Article
- Keywords:
avian reovirus;
σC gene;
pMD19T-σC;
pET32a-σC
- From:
Chinese Journal of Veterinary Science
2009;29(7):836-840
- CountryChina
- 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.
