KGF-2与PRO2605蛋白相互作用的初步研究

目的：利用酵母双杂交系统筛选KGF-2的相互作用蛋白，为阐明其分子作用机制提供有益线索。方法：通过聚合酶链反应从人胎肝cDNA文库中钓取KGF-2 cDNA，构建诱饵蛋白载体pAS2-1-KGF-2，并对其自身转录激活活性进行鉴定，利用酵母双杂交系统筛选人胎肝cDNA文库，挑选双阳性克隆。将KGF-2和候选蛋白分别克隆至哺乳动物细胞双杂交的BD、AD质粒中，共同转染COS-7细胞，通过CAT分析验证KGF-2和候选蛋白之间的相互作用。结果：VDNA序列分析和同源检索显示所获候选蛋白为PR02605。以KGF-2和PRO2605 cDNA共转化酵母宿主Y190后可激活报道基因，但共转染COS-7细胞后，CAT分析结果阴性。结论：KGF-2和PRO2605在酵母体内存在相互作用，但在COS-7细胞中未能检测到两者的相互作用。

Interaction of KGF-2 with PRO2605:a preliminary study

Objective:To explore the protein interactive with ker at inocyte growth factor-2 (KGF-2). Methods:The cDNA en coding KGF-2 was isolated from human fetal liver cDNA library and subsequently cloned into the bait protein plasmid pAS2-1.The positive clones were screened o ut of human fetal liver cDNA library by yeast two-hybrid system. KGF-2 and the candidate protein were separately cloned into BD and AD plasmid of mammalian tw o-hybrid systim and cotransfected into COS-7 cell line. The interaction betwee n KGF-2 and the candidate protein was identified by CAT assay. Resul ts: After sequence analysis and homology comparison, the candidate p r otein was identified as PRO2605. KGF-2 and PRO2605 could activate the transcrip tion of report gene. Conclusion: KGF-2 could interact wit h PRO2605 in yeast host Y190,but the interaction was not detected in COS-7 cel l line.