大麦条纹花叶病毒中国株编码βC蛋白可以自身相互作用

以分别含有BSMV-CH基因组3个组分ds-cDNA全长的pMD-α、pMD-β和pMD-γ质粒为模板,PCR扩增BSMV-CH编码各蛋白基因,克隆至酵母双杂交载体,检测各蛋白是否存在自身激活特性,用酵母双杂交分析BSMV-CH编码蛋白之间的相互作用情况,测定β-半乳糖苷酶活性确认蛋白在酵母体内的作用,Western-blot分析蛋白的体外相互作用。结果表明成功克隆到BSMV-CH编码蛋白基因,并正确克隆至酵母双杂交载体,酵母双杂交检测未发现BSMV-CH编码蛋白之间的相互作用,但确定BSMV-CH编码的βC蛋白可以自身相互作用,体外可以单体或二聚体形式存在。

Interaction of βC protein of Barley stripe mosaic virus China strain (BSMV-CH) with itself

The protein genes of BSMV-CH were amplified by PCR from the recombined plasmids pMD-α, pMD-β and pMD-γ, which contain 3 segments of the full-length ds-cDNA of BSMV-CH respectively.The obtained fragments were cloned into vectors of yeast two-hybrid system for analyzing the protein-protein inte-ractions between BSMV-CH proteins after detecting the self-activity of each protein.The β-galacatosidase activity was tested to ensure the protein-protein interaction in the yeast cells.The protein-protein interactions in vitro were detected by western blot.The results showed that the BSMV-CH encoded protein genes were amplified and cloned into the vector of yeast two-hybrid system.No protein-protein interaction was found in yeast two-hybrid system, but the self-interaction of BSMV-CH βC protein was detected, which could keep dimer or monomer in vitro.