LRIG1基因过表达载体的构建、鉴定及在食管鳞癌细胞株Eca109中的表达  

Construction,identification of LRIG1 over-expression vector and expression in esophageal cancer strain Eca109

作　　者：吉别克.瓦提别克 李惠武[2] 喻亮 路晨菲 范志勤[2] 李颖[1] 李卉[5] 刘玲[1] JIBIEKE·Watibieke;LI Hui-wu;YU Lian;LU Chen-fei;FAN Zhi-qin;LI Ying;LI Hui;LIU Ling(Preclinical College;Affiliated Tumor Hospital;Public Healthy College;Clinical Medical College;Central Laboratory, Xinjiang Medical University, Urumqi Xinjiang, 830011, China)

机构地区：[1]新疆医科大学基础医学院,新疆乌鲁木齐830011 [2]新疆医科大学附属肿瘤医院,新疆乌鲁木齐830011 [3]新疆医科大学公共卫生学院,新疆乌鲁木齐830011 [4]新疆医科大学临床医学院,新疆乌鲁木齐830011 [5]新疆医科大学中心试验室,新疆乌鲁木齐830011

出　　处：《职业与健康》2018年第5期611-616,共6页Occupation and Health

基　　金：国家自然科学基金项目(81460419);新疆维吾尔自治区大学生创新性实验计划项目(201610760055);新疆医科大学大学生创新性实验计划项目(CX2016055)

摘　　要：目的构建LRIG1基因过表达载体,并进行鉴定。方法采用PCR方法从食管癌的LRIG1c DNA表达阳性的标本中扩增出LRIG1基因,经用Xho I和Kpn I-HF双酶切后,与过表达载体GV219相连后转入E.coli DH5α。结果 DNA测序证明获得了包含ORF全长的LRIG1基因,并成功克隆入过表达载体GV219中。经转染Eca109细胞株后,应用蛋白免疫荧光技术检测过表达的LRIG1。结论构建过表达载体成功,为进一步探讨LRIG1基因在食管癌中的作用奠定了基础。[Objective]To construct and identify a lentiviral vector for over-expression of LRIG1 gene.[Methods]PCR was used to clone LRIG1 gene with esophageal cancer c DNA as a template.After digesting with endonuclease enzyme Eco RI and Kpn I-HF,LRIG1 gene was linked with eukaryotic expression vector GV219 and transferred into E.coli DH5α.[Results]DNA sequence which included ORF was obtained,and which was successfully linked with eukaryotic expression vector GV219.After transfection with Eca109 strain,over-expression LRIG1 gene was detected by protein immunofluorescence technique.[Conclusion]The lentiviral vector over-expressing LRIG1 gene is successfully constructed,which lays the foundation for further study of the role of LRIG1 gene in esophageal cancer.

关 键 词：LRIG1表达载体 构建 鉴定 

分 类 号：R734.2[医药卫生—肿瘤]