小鼠整合素α_4基因克隆、序列分析及突变修复  

Molecular cloning, sequence analysis of mouse integrin α_4 gene and repairing of its mutation

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：《第三军医大学学报》2007年第22期2131-2134,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30400408)~~

摘　　要：目的克隆小鼠整合素α4基因cDNA,进行分析整合素α4基因序列,对造成氨基酸突变的碱基进行定点突变修复。方法从小鼠小肠peyer’s结获得α4基因cDNA,采用RT-PCR方法,克隆到pMD18-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α4基因cDNA为3099bp,编码1032个氨基酸,与GenBank中公布的序列比较,有12个碱基发生突变,其中6个碱基造成6个氨基酸发生改变,对发生突变的6个碱基进行定点突变修复。结论获得小鼠整合素α4基因的克隆。Objective To clone and analyze the full-length cDNA of mouse integrin α4, and repair the mutation sensible locei that caused the change of amino acids. Methods The cDNA of α4 gene was amplified by RT-PCR using the total RNA extracted from mouse small intestine peyer＇ s patch. The PCR product was inserted into pMD19-T vector and then transformed into E. coli JM109. The positive recombinant clone was analyzed by restriction endonuclease and DNA sequencing. The mutation of α4 cDNA that caused the change of amino acids was repaired. Results The cDNA of mouse α4 had a length of 3 099 bp, and encoded a product of 1 032 amino acids. There were 12 bases pairs mutation of α4 gene and the 6 base pairs causing the change of amino acids was repaired. Conclusion The cDNA of mouse α4 is cloned successfully.

关 键 词：α4基因 基因克隆 RT-PCR 突变修复 

分 类 号：R322.45[医药卫生—人体解剖和组织胚胎学] R394.33[医药卫生—基础医学]