摘要
目的应用酵母双杂交技术研究能够与维甲酸诱导的G基因 (RIG -G)发生作用的蛋白 ,以了解其功能。 方法应用酵母双杂交技术 ,以RIG -G为诱饵 ,筛选人类骨髓cDNA文库 ,寻找能与之相互作用的蛋白。利用生物信息学 ,一对一酵母回复性杂交 ,β -GAL实验等提供的信息 ,筛除假阳性克隆 ,确定阳性克隆。 结果经过酵母双杂交共获得 10 9个克隆 ,经过验证 ,有 2 4个阳性克隆 ,它们分别是细胞的信号传导蛋白、细胞骨架蛋白、激酶等。 结论为了研究诸如RIG -G新基因的功能 ,酵母双杂交技术是可靠的分子生物学技术。
Objective The aim of this study is to research proteins interacting with RIG-G gene by yeast-two-hybrid technique. Methods Using RIG-G as bait, a human bone marrow cD NA library was screened and the proteins interacting with RIG-G were searched. The false positive clones were discarded by one to one yeast-two-hybrid, β-gal assay, and bioinformatics methods. Results We obtained 24 true positive clones from 109 yeast clones after verification. These proteins play important roles in cell proliferation, cell differentiation, signal transduction and cell skeleton. Conclusion The yeast-two-hybrid technique is a reliable method for studying the function of a new gene such as RIG-G.
出处
《上海第二医科大学学报》
CSCD
2001年第5期385-389,400,共6页
Acta Universitatis Medicinalis Secondae Shanghai
基金
上海市青年科技启明星计划 (99QB1 4 0 2 4 )
中法高科技先进项目 (PRA) (B98- 0 2 )
上海市教委科技发展基金项目 (98B1 2 )
国家自然科学基金重大项目 (3999342 0 )
国家自然科学基金面上项目 (2 0 0 1 -2 0 0 3)
上海血液学研究所胡应洲基金资助项目
