›› 2011, Vol. 38 ›› Issue (7): 76-79.

• 生物技术 • Previous Articles     Next Articles

Cloning and Construction of Prokaryotic Expression Vector of B2L Gene of Orf Virus in Guizhou

XIANG Zhi-long1, ZHUO Jian-hua3, CHENG Zhen-tao1,2, OU De-yuan1,2, XIAN Si-mei1,2, YIN Chuan-bao1, HUANG Lu1, HE Cai-hong1   

  1. 1. College of Animal Science, Guizhou University,Guiyang 550025,China;2. Insitute of Animal Disease of Guizhou Province,Guiyang 550025,China;3. The Bureau of Livestock and Veterinary of Jingshan Country,Jingshan 431800,China
  • Received:2010-11-30 Revised:1900-01-01 Online:2011-07-20 Published:2011-07-20

Abstract: To identify suspected goats orf case and construction of prokaryotic expression vector of B2L gene of orf virus (ORFV),clinical and suspect samples of orf could be amplified by a pair of specific primers,then the structural protein B2L gene of orf virus was amplified by PCR method using specific primers.The B2L gene was ligated with pMD18-T vector and the sequencing results showed that the full-length of the inserted gene fragment was 1137 kb. Cutting the target fragment,which was cloned into pET-32a vector and construction of prokaryotic expression vector pET-32a-B2L,the recombinant plasimid was identified by restriction enzyme and PCR,sequence analysis indicated that to be successful construction of pET-32a-B2L expression vector and it has laid a good foundation for the expression and genetic engineering vaccine research.

Key words: ORFV; clone; prokaryotic expression vector

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