摘要
目的构建ezrin基因RNA干扰慢病毒载体并建立ezrin基因稳定干扰的人胃癌SGC-7901细胞系,为ezrin基因的功能研究奠定基础。方法设计ezrin基因特异性RNA干扰靶序列,与PHR-SIN-CSGW-H1a载体定向连接,构建PHR-SIN-CSGW-H1a真核入门表达载体。通过与psPAX2和PMD2.G载体进行重组,获得ezrin基因真核表达重组慢病毒表达载体。利用包装细胞293FT获得重组的慢病毒,感染人胃癌SGC-7901细胞。结果成功构建PHR-Eai-psPAX2-PMD2.G真核表达慢病毒干扰载体并获得相应的慢病毒,病毒悬液的滴度>1×107U/L。RT-PCR、Western blot结果表明,ezrin基因mRNA及蛋白表达显著降低。结论成功构建人ezrin基因特异性的重组慢病毒干扰载体,获得ezrin基因稳定干扰的人胃癌SGC-7901细胞系。
Objective To construct a lentiviral vector for RNA interference of ezrin gene and to establish a human gastric cancer cell line with ezrin gene interference.Methods Different short hairpin RNAs targeting ezrin gene-specific sequence was designed to link with PHR-SIN-CSGW-H1a vector,then constructed the entry lentiviral vector.A recombinant lentiviral vector was obtained using the PHR-SIN-CSGW-H1a of psPAX2 and PMD2.G vector.The recombinant lentiviral vectors were harvested from 293FT cells and were used to ...
出处
《胃肠病学和肝病学杂志》
CAS
2009年第11期1007-1011,共5页
Chinese Journal of Gastroenterology and Hepatology
