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E—cadherin启动子调控的慢病毒荧光报告载体的构建
引用本文:何倩,杨耿兵,姚超,钱程. E—cadherin启动子调控的慢病毒荧光报告载体的构建[J]. 浙江理工大学学报, 2012, 0(6): 843-846,862
作者姓名:何倩  杨耿兵  姚超  钱程
作者单位:浙江理工大学生命科学学院,杭州310018
基金项目:国家自然科学基金重大项目(81090423)
摘    要:E钙黏素(Dcadherin)的缺失是上皮一间质转化(EMT)的重要标志,而EMT可促进肿瘤细胞的侵润及转移。为进一步研究E-cadherin与EMT的关系,研究构建了N-cadherin启动予驱动的黄色荧光蛋白慢病毒表达载体,构建的载体经酶切鉴定正确;采用磷酸钙法包装出慢病毒,取病毒上清感染人肝癌PLC/PRF/5细胞株,通过荧光观察和qRT-PCR实验结果显示E-cadherin在靶细胞中稳定高表达。该载体的构建为深入研究E-cadherin相关的EMT现象提供了良好的工具。

关 键 词:E-cadherin  慢病毒载体  黄色荧光蛋白

Construction of Lentiviral-based System with Fluorescent Reporter Gene Regulated by E-cadherin Promoter
HE Qian,YANG Geng-bing,YAO Chao,QIAN Cheng. Construction of Lentiviral-based System with Fluorescent Reporter Gene Regulated by E-cadherin Promoter[J]. Journal of Zhejiang Sci-tech University, 2012, 0(6): 843-846,862
Authors:HE Qian  YANG Geng-bing  YAO Chao  QIAN Cheng
Affiliation:(School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China)
Abstract:The loss of E-cadherin is a significant marker of epithelial-mesenchymal transition (EMT), which promotes the progression of tumor invasion and metastasis. To further examine the relationship be- tween E-eadherin and EMT, the lentiviral vector with yellow fluorescent protein reporter gene driven by E- cadherin promoter is successfully constructed. The supernatant of virus-producing cells transfected by cal- cium phosphate is then used to transfect PLC/PRF/5 cells. The fluorescent microscopy and qRT-PCR re- sults show that the E-cadherin gene is expressed highly and stably in the target cells. This work is expec- ted to provide a high-quality transfection vector/or further research on the relevant functions of the E-cad- herin gene and EMT.
Keywords:E-cadherin  lentiviral vector  yellow fluorescent protein
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