Cloning of the E2 gene of classical swine fever virus and construction of a baculovirus expression vector

Ji XinCheng; Chen Jie; Zhang YanMing; Wu FaXing; Li XiaoCheng; Huang BaoXu; Zhang YanXia; Xu XinGang

Chinese Journal of Veterinary Science and Technology 34(1): 41-44

2004


Accession: 004076683

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Abstract
The E2 gene of classical swine fever virus (CSFV) virulent strain from Shaanxi Province of China was amplified by RT-PCR. The E2 fragment was cloned into pMD 18-T vector. The cDNA fragments of the E2 gene were sequenced. Based on the amino acid sequences of the China strain, Brescia strain and Alfort strain, the amino acid sequence encoded by the E2 gene was deduced. The nucleotide sequence homologies of the prevalent virulent strain with China, Brescia and Alfort strains were 81.9, 83.4 and 83.5%, respectively. The E2 gene cDNA was subcloned into baculovirus transfer vector and the recombinant baculovirus vector pFBHT-E2 was constructed successfully. It was then transferred into E. coli DH10Bac and cultured in LB plate. The white bacterial colonies were positive recombinant bacmid named rBacmid-E2. The results lay down the theoretical and practical foundations for the expression of E2 gene in insect cells.