国家自然科学基金资助项目(30271480).
This work was supported by a grant from The National Natural Sciences Foundation of China (30271480).
通过聚合酶链式反应从人胎肝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.DNA序列分析和同源检索显示,所获侯选蛋白为人核糖体蛋白L22(RPL22).将KGF-2和侯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分析验证了KGF-2和侯选蛋白之间的相互作用.为阐明KGF-2作用的分子机制提供有益线索.
The cDNA encoding keratinocyte growth factor-2 was isolated from human fetal liver cDNA library and subsequently cloned into the bait protein plasmid pAS2-1.The positive clone was obtained from human fetal liver cDNA library screened by yeast two-hybrid system. After sequence analysis and homology comparison, the candidate protein was identified as human ribosomal protein L22.KGF-2 and the candidate proteins were separately cloned into BD and AD plasmid of mammalian two-hybrid assay and cotransfected into COS-7 cell line. The interaction between KGF-2 and the candidate protein was identified by CAT assay.
白秉学,徐东刚,王金凤,彭善云,邹民吉,范明.角质细胞生长因子-2与核糖体蛋白L22相互作用的发现及在哺乳动物细胞中的验证[J].生物化学与生物物理进展,2004,31(10):936-940
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