Cloning And Sequence Analysis On E2 Gene Of Different Cell Strains Infected By Hog Cholera Virus Lapinized Chinese Strain

In this study, Hog cholera Virus lapinized chinese strain can grow in Bovine Testicle Cell and Swine Testis Cell, and detect growth characteristics, HCLV multiplication and no others viruses. The further study of genetic relationship between the major protective antigen encoding regions of E2 genes of ST cell strain and BT cell strain was done by molecular biology heredity methods.The nucleic acids of Hog cholera viruses conducted in the research were extracted from the viruses propagated in ST cell and BT cell. E2 Gene of different cell strains infected by HCLV were amplified by RT-PCR (Reverse transcription-polymerase chain reaction). The cDNAs were cloned into pMD18-T vector and identified. The sequencing products were purified and sequenced. Finally, using DNAStar software was used for data analysis, sequence translation, phylogenetic tree drawing and molecular evolution analysis.The results indicated that HCLV can multiplicated in Bovine Testicle Cell and Swine Testis Cell,and cell's stabilization growth characteristics, and no exogenous viruses.Their nucleotide sequences were determined and the amino acid sequences were deduced. Compared with the corresponding region of AY663656-C81, their homologies among seven strains were very higher. That is, their nucleotide sequence homologies and amino acid homologies were 99.0%～99.4%,98.7%～99.7% respectively. And the nucleotide sequence homologies and amino acid homologies were 99.0%～99.4%, 98.7%～99.7% respectively, compared with Z46258-C. The phylogenetic trees revealed that E2 genes of ST cell strain and BT cell strain, AY663656-C81 and Z46258-C conducted in this study belonged to the same group, derived from AF092448-Shimen.Furthermore, the variation analysis of amino-acid in the major protective antigen encoding regions of E2 genes of all strains, showed that E2 genes of ST cell strain be none mutation in A,B,C and D antigen regions, While there is subtle difference in E2 genes of BT cell strain's A,B,C and D antigen regions. The research confirmed that E2 genes of ST cell strain and BT cell strain infected by HCLV were genetically stable.In conclusion, this study elucidated HCLV multiplication be cultured in Bovine Testicle Cell and Swine Testis Cell,and E2 genes of ST cell and BT cell strain be none or subtle difference in A,B,C and D antigen regions by molecular heredity evolution methods. The research ensured vaccine virus in ST cell and BT cell favourable suitability and antigenicity. Of particular significance, these would provide the theory support for the possibility and development trend of the classical swine fever vaccine in ideal immunological effect.