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Cloning, Expression and Identification of Matrix Protein Gene of Wheat Rosette Stunt Virus (WRSV)
1Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. gongzx@sunm.shcnc.ac.cn
Abstract:
cDNA fragment located downstream of the WRSV NS protein gene was sequenced. Following an 80 nt untranslated region at the 5 'terminus, there is a 453 nt open reading frame(ORF) encoding a 17 kD protein, which was allowed to be expressed in E. coli using bacterial expression vector pGEX-3X and produced a 43 kD fusion protein. The result of Western blot showed that the fusion protein was able to react strongly with antibody raised against the purified WRSV particles. According to the similarities between the gene organizations of VSV and WRSV and between the molecular weights of deduced and expressed proteins, as well as the viral structural M protein, and to the result of Western blot, the 453 nt ORF was identified as WRSV M protein gene.
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