Cloning And Analysis The Genome Sequence Of Barley Stripe Mosaic Virus China Strain And Studies On Its Protein-protein Iteraction

Barley stripe mosaic virus (BSMV) is the type member of the Hordeivirus. BSMV had been distributed all over the world by seed transmission. It had prevailed in many regions of Xinjiang province and was spread to many areas of China, which caused great loss in agriculture production. Therefore, it would be important and useful to do research on the diversity of BSMV strains, the interaction among the proteins of BSMV and the interaction between the proteins of BSMV and that of its hosts.Extensive studies had been done on the biologic and molecular character of BSMV. The complete genome sequence of CV17, ND18 and CV42 had been published one after another since the complete genome sequence of the Type strain were first reported by Gustafson in 1986. However, no complete genome sequence was reported after the 5`cap structure , the 3` and 5`sequence of BSMV-XJ were reported by Xuexian Peng and his colleague in China in 1989. In 2004, we found barley which may be infected by BSMV, and collected the infected barley leaves to identify the pathogeny in the leaves by biological and serological methods in our laboratory. Then, we amplified the cDNA of the complete genome of the BSMV isolated in our lab with primers designed on the base of the published complete genome sequence of BSMV. The cDNA were cloned into the pMD-18 vector, and their sequences were sequenced and analyzed by comparing.We amplified the coat protein (CP) of BSMV from the c DNA of BSMV RNAβ,and constructed the GST-CP fusion vector to expressing the GST-CP in E.ciol BL21.After optimizing the expression condition, we expressed the GST-CP in E.ciol BL21, and purified the GST-CP to immunized the rabbit for obtaining the antiserum.Yeast two hybrid system (YTHS), established by Fields and his colleague, is an sensitive technology to identify the protein-protein interaction in yeast cell. It could be rapid and sensitive in detecting the interaction in vivo, and obtaining the gene of the target protein. YTHS has been extensively used in studying the control of cell cycle and gene, conduction of transmembrance signal and drug design, et al. In recent year, YTHS has also be used in study the molecular structure and assemblage of plant virus, replication of nucleic acid, control of gene expression, molecular mechanism of aphid transmission and the map of the protein interaction, et al. we amplified the 7 proteins encoded by BSMV from the cDNA of its complete genome and cloned them into pGBKT7 and pGADT7 to detect the protein-protein...