人整合素β3亚基编码基因真核表达载体的构建  

Consttuction of eukaryotic expression vector harboring human integrin beta 3 gene

作　　者：牟丹蕾[1] 白雪帆[1] 李光玉[1] 潘蕾[1] 黄长形[1] 杨为松[1] 

机构地区：[1]第四军医大学唐都医院全军感染病防治中心,西安市710038

出　　处：《热带病与寄生虫学》2003年第2期72-75,共4页Journal of Tropical Diseases and Parasitology

摘　　要：目的构建人整合素β3亚基金读码框(ORF)基因真核表达载体,为探讨整合索β3作为汉坦病毒(Hantavirus,HV)受体的特异性奠定基础。方法根据已公布的序列设计引物,用PCR方法从原核质粒中扩增出人整合素分子β3亚基ORF基因,应用TA克隆将其插入pcDNA3.1/V5-His-TOPO载体中,采用酶切和PCR鉴定,选取初筛正向插入的阳性克隆进行测序。结果序列分析表明与基因库中已发布的人整合素β3亚基ORF核苷酸序列基本一致,成功构建了pcDNA3.1-β3真核表达载体。结论上述结果表明,真核表达载体pcDNA3.1-β3已被成功构建,并为进一步研究汉坦病毒的病原学及生物学特征提供了基础。Objective To study the cellular entry of hantavirus we constructed eukaryotic expression vec- tors harboring Homo sapiens integrin β3 gene. Methods The 2.3kb ORF gene of human integrin β3 subunit was amplified from the plasmid containing cDNA of human integrin β3 by PCR. Then the PCR products with the blunt-ends were directly cloned into eukaryotic expression vector pcDNA 3.1/V5-His TOPO by TA Cloning. Re- sults It shows that The integrin gene was inserted into the plasmid vector by restriction enzyme analysis and PCR, and the sequence is identical with the reported human integrin β3 ORF gene. Conclusion The results above sug- gested that the eukaryotic expression vectors, pcDNA3.1-β3, has been successfully constructed and it lies a basis for the further study of hantavirus pathogenicity and biological characteristics.

关 键 词：整合素 β3亚基 编码基因 真核表达 载体 CDNA克隆 序列测定 聚合酶链反应 

分 类 号：R394[医药卫生—医学遗传学]