小鼠整合素β_7基因克隆、序列分析及突变修复  

Molecular cloning,sequence analysising and repairing the mutation of mouse integrin β_7 gene

作　　者：王永明[1] 胡燕[1] 黎海芪[1] 

机构地区：[1]重庆医科大学附属儿童医院儿童保健科,重庆400014

出　　处：《重庆医科大学学报》2007年第10期1027-1031,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金(30400408)。

摘　　要：目的:克隆小鼠整合素β7基因cDNA,同时对其序列进行分析,对造成氨基酸突变的碱基进行定点突变修复。方法:采用RT-PCR的方法,从小鼠小肠PP结获得β7基因cDNA,克隆到pMD19-T载体,选择阳性克隆进行酶切鉴定和序列测定,对造成氨基酸突变的碱基进行定点突变修复。结果:扩增得到的β7基因cDNA为2418bp,编码806个氨基酸,与GenBank中公布的序列比较,有10个碱基发生突变,其中造成氨基酸发生改变的有5个碱基,涉及3个氨基酸,对发生突变的5个碱基进行定点突变修复。结论:获得小鼠整合素β7基因的克隆,为进一步研究其生物学功能奠定了良好基础。Objective：To clone and analyze a full-length cDNA encoding mouse integrin β7,and repair the mutation of β7 eDNA that caused the change of amino acids. Methods：The eDNA of β7 gene was amplified by RT-PCR using the total RNA as extracted from mouse small intestine Peyer＇s patch. The PCR product was inserted into pMD19-T vector and then transformed E.eoli JM109. The positive recombinant clone was analyzed by restriction endonuelease and DNA sequencing. The mutation of β7 eDNA that caused the change of amino acids was repaired. Results：The eDNA of mouse β7 has an complete open reading frame with a length of 2418 bp,whieh encodes a product of 806 amino acid,and has 10 base pairs mutation of β7 gene and 5 base pairs mutation that caused the change of amino acids was repaired. Conclusion：The eDNA of mouse β7was cloned successfully,which posed a basis for further researching on its biological function.

关 键 词：β7基因 基因克隆 RT-PCR 突变修复 

分 类 号：R72[医药卫生—儿科]