2009•Guizhou nongye kexueRequires access
Cloning and prokaryotic expression of full VP4 gene of group A bovine rotavirus.
Xiaowei Nan, Yang ShaoHua, Yun‐Dong Gao, Wagn ChangFa, Hongjun Yang, Xiao Liu, Jifeng Zhong, Shen Zhi-yi, Hongbin He
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Abstract
In order to construct the prokaryotic expression vector pET32a-VP4,total RNA were extracted form MA104 cell infected by bovine rotavirus G6 strain CHLY.The open reading frame of VP4(2331bp) was amplified by RT-PCR and cloned into pEASY-T3 vector by using T-A cloning technique to get cloning vector pEASY-T3-VP4.After,the VP4 gene was subcloned into the expression vector pET32a and transformed into E.coli BL21(DE3).SDS-PAGE showed that the bacteria induced by IPTG successfully expressed,and approximately 108KD exogenous protein was observed on the SDS-PAGE.The with of VP4 was successfully constructed which is expected to lay a foundation for further studies of the subunit vaccine and DNA vaccine to prevent bovine rotavirus.