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.
作者
吉别克.瓦提别克
李惠武
喻亮
路晨菲
范志勤
李颖
李卉
刘玲
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)
出处
《职业与健康》
CAS
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)
关键词
LRIG1表达载体
构建
鉴定
LRIG1 expression vector
Construction
Identification
