【摘要】 目的观察结核分枝杆菌Rv1246c和Rv1247c假想蛋白间的相互作用。方法采用PCR技术克隆结核分枝杆菌Rv1246c和Rv1247c假想蛋白基因,构建酵母双杂交载体质粒pGBKT7-Rv1247c和pGADT7-Rv1246c,经酶切分析和DNA测序证实重组质粒构建成功后,采用醋酸锂法顺序转染酵母菌AH109,检测β半乳糖苷酶活性。结果共转染GBKT7-Rv1247c和pGADT7-Rv1246c的酵母菌AH109可以在SD/-Ade/-His/-Leu/-Trp营养缺陷培养基上生长,β半乳糖苷酶活性实验阳性。结论利用酵母双杂交系统证实结核分枝杆菌Rv1246c与Rv1247c假想蛋白可以相互作用。更多还原

【Abstract】 Objective To investigate the protein-protein interaction between hypothetical protein Rv1246c and Rv1247c of Mycobacterium tuberculosis. Methods By PCR technique, the complete open-reading frame sequences of Rv1246c and Rv1247c gene were amplified from the M.tuberculosis H37Rv genomic DNA as template. The PCR-amplified cDNAs of Rv1247c and Rv1246c gene were subcloned into pGBKT7 and pGADT7-Rec vector respectively for constructing recombinant plasmids pGBKT7-Rv1247c and pGADT7-Rv1246c. After verified by restriction endonuclease digestion and DNA sequence determination, the recombinant vectors were used to transform the yeast cell AH109 by lithium acetate method. Results The yeast cells co-transformed with pGBKT7-Rv1247c and pGADT7-Rv1246c grew on SD/-Ade/-His/-Leu/-Trp plates, and the β-galactosidase activity assays showed the positive signal. Conclusion The hypothetical protein Rv1246c and Rv1247c could interact with each other in yeast cells.更多还原