Cloning And Identification Of The Antiapoptosis Virulent Factor Gene Of Goatpox Virus In Guizhou

Viruses used in this study were isolated from Guizhou in China .The control of apoptosis by goatpox viruses was studied during goatpox viruses infecte the permissive cells in vitro, then an important factor interrelated apoptosis---Antiapoptosis virulence factor of goatpox virus was studied further.The structure of antiapoptosis virulence factor gene, the mRNA dynamics changes of antiapoptosis virulence factor gene in the process of permissive cells infectd viruses ,the relationship between the expression of antiapoptosis virulence factor gene and apoptosis were studied and analyed in this paper.The pathological changes of the Vero cells infected goatpox viruses were observed by the inverted microscope.The rate of cells death was monitored by typan blue exclusion staining .The apoptosis induced by goatpox virus was observed by AO/EB fluorescence staining and .DNA was extracted from the Vero cells with treatment Goatpox viruses. The 531 nucleotide acids were amplificated by PCR. The comparability of the antiapoptosis virulence factor gene and the protein encoded by the antiapoptosis virulence factor gene was analysed respectively on line. The amino acid composition , physicochemical property, hydrophobicity and transmembrane domain and secondary structure of the protein encoded were analysed by biosoftware .The expression of antiapoptosis virulence factor gene mRAN was detectd by RT-PCR technique.In Vero cells with treatment of viruses for 36 hours, the shape of cells has become round , there were wire drawings between cells , and with treatment of viruses for 48 hours, the most cells began falling from the wall, which were observed by inverted miscroscope, when cells with treatment of viruses for 36 hours , the rate of cells death reached 80 percentage, and with treatment of viruses for 48 hours , the rate of cells death reached 100 percentage. The results showed that there were no obviously DNA ladders, the mRAN expression of antiapoptosis virulence factor gene is different .Before the cells were infected by viruses for 24 houres, the mRAN expression is increasing following the time infected increasing, and at the age of 24 hour, it reach the maximum. ,and then was decreasing .It suggested that the protein encoded should have an important function of regulation between goatpox virus and apoptosis, it maybe control the time of cell disruption.The study will have an important significance in illustrating the pathogenesis mechanism of goatpox virus,antiviral remedy filtration and preventation of the goatpox in Guizhou province.