牛整合素β1亚基在COS-7细胞中的真核表达

摘要

The integrin is formed by 18 kinds of alpha subunit and 8 kinds of beta subunit on the cell surface. Integrin beta β1 gene is a major member of the adhesion molecule family,and plays an important role in cells amplification, migration and survival. In this study, specific primers targeting at the bovine integrin β1 gene were used to amplify the gene and then cloned the eukaryote expres-sion vector pEYFP-C1 to construct for recombinant plasmids pEYFP-C1-β1, which was identified by PCR methods and enzyme di-gestion. Then the positive plasmids and liposome were co-transfection into COS-7 cells, after 48 h excellent fluorescent signals can be observed at COS-7 cells. All the results demonstrated that eukaryote expression of the bovine integrin β1 gene in COS-7 cells were successed with good biological activity, which lays good foundation for the research of bovine integrin β1 gene biological activity.%整合素是由18种α亚基和8种β亚基在细胞表面形成的异型二聚体。研究发现，整合素β1亚基是粘附分子家族主要成员，对多种细胞的存活、扩增、迁移有重要作用。本研究通过设计针对牛整合素β1亚基的特异性引物，将其克隆到真核表达载体pEYFP-C1上，构建pEYFP-C1-β1重组表达质粒，经PCR和酶切鉴定符合试验预期的阳性重组质粒与脂质体按照1：1.5比例共转染COS-7细胞，48 h后可见阳性重组质粒共转染COS-7细胞出现荧光信号。结果表明，本研究成功构建出具有生物学活性的牛整合素β1亚基，为研究牛整合素β1亚基的生物学功能奠定基础。