[结核分枝杆菌ESAT-6与CFP-10之间的蛋白质-蛋白质相互作用]
[Protein-protein interaction between ESAT-6 and CFP-10 of mycobacterium tuberculosis].
作者信息
Li Yan, Bao Lang, Zhang Hui-dong, Wang Xiao-ying, Zhu Hai-long, Li Ya-sha
机构信息
Research Unit of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
出版信息
Sichuan Da Xue Xue Bao Yi Xue Ban. 2006 May;37(3):349-52.
OBJECTIVE
To investigate the protein-protein interaction between ESAT-6 and CFP-10 of mycobacterium tuberculosis.
METHODS
ESAT-6 gene and CFP-10 gene were amplified from mycobacterium tuberculosis genome DNA. The ESAT-6 gene was subcloned into pGBKT7 and the CFP-10 gene was subcloned into pGADT7-Rec. After being verified with restriction endonuclease digestion and DNA sequencing, the recombinant vectors were transformed into yeast cell AH109 by lithium acetate method.
RESULTS
The yeast cells co-transformed with pGBKT7-ESAT-6 and pGADT7-CFP-10 grew on SD/-Ade/-His/-Leu/-Trp plates, and beta-galactosidase activity assays showed positive results.
CONCLUSION
ESAT-6 and CFP-10 protein could interact with each other in yeast cells.
目的
研究结核分枝杆菌ESAT-6与CFP-10之间的蛋白质-蛋白质相互作用。
方法
从结核分枝杆菌基因组DNA中扩增ESAT-6基因和CFP-10基因。将ESAT-6基因亚克隆到pGBKT7中，将CFP-10基因亚克隆到pGADT7-Rec中。经限制性内切酶消化和DNA测序验证后，通过醋酸锂法将重组载体转化到酵母细胞AH109中。
结果
共转化pGBKT7-ESAT-6和pGADT7-CFP-10的酵母细胞在SD/-Ade/-His/-Leu/-Trp平板上生长，β-半乳糖苷酶活性测定显示阳性结果。
结论
ESAT-6和CFP-10蛋白在酵母细胞中可相互作用。
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