【摘要】 食管癌的形成与多种基因的异常表达有关,哈萨克族食管癌的发病又有其独特的机制。克隆并研究早期相关基因的启动子区CpG岛可为进一步揭示其发病机制奠定基础。本研究构建Survivin基因启动子区CpG岛真核表达载体,探讨其甲基化在新疆哈萨克族食管癌中的作用。通过Primer5.0设计软件设计引物,采用常规PCR法从新疆哈萨克族食管癌患者的基因组DNA中扩增出Survivin基因启动子区CpG岛,CpG岛与真核表达载体pCAT3promoter经HindⅢ单酶切后,连接试剂盒将二者连接并转导入甲基化酶缺陷的大肠杆菌E.coli ER1793中扩增。DNA测序证明获得Survivin基因启动子区CpG岛,并成功克隆入真核表达载体pCAT3promoter中。更多还原

【Abstract】 There are many kinds of genes’ abnormal expressions participating in the form of esophageal cancer. Hazak’s esophageal cancer has a special mechanism. Cloning and studying the early related genes’ promoter CpG island would provide a basis for revealing the mechanism. The Survivin promoter CpG island in Hazak’s esophageal cancer in Eukaryotic Expression Vector is constructed in this study for identifying the methylation function of the CpG island on Hazak’s esophageal cancer. By Primer 5.0 software, the primer is designed, and the Survivin promoter CpG island is cloned with PCR from Hazak’s esophageal cancer in genome DNA, after cutting by restrictive endonuclease enzyme HindIII, linking the fragment to eukaryotic expression vector pCAT3 promoter and transferring into E. coli ER1793. After the DNA sequence, the fragment is successfully linked with pCAT3 promoter eukaryotic expression vector. As a result, the eukaryotic expression vector is constructed, which may help studies of the effects of Survivin promoter methylation on Hazak’s esophageal cancer.更多还原