山东医药2016，Vol.56Issue(21)：1-3,3.DOI:10.3969/j.issn.1002-266X.2016.21.001
Eμ-VH 启动子调控 c-Myc 表达慢病毒载体的构建及其体外功能
Construction and function of a lentiviral vector harboring c-Myc gene under the control of Eμ-VH promoter
摘要
Abstract
Objective To construct a lentiviral vector harboring c-Myc gene under the control of human B lymphocyte-specific Eμ-VH promoter (pEMIR),and to investigate its function.Methods Firstly,taking pLV-c-Myc-IRES-EGFP as a template,after PCR amplification and enzyme digestion,we got pEVCHR.Secondly,taking plasmid pEVCHR as a tem-plate,after PCR amplification and enzyme digestion,we got a lentiviral vector pEMIR.pEMIR was transiently transfected into human embryonic kidney cells 293T and human diffuse large B-cell lymphoma cell line (OCL-LY3 or OCL-LY10). The expression of enhanced green fluorescent protein (EGFP)and mono-red fluorescence protein (mRFP)was detected under inverted fluorescence microscope,and the expression of c-Myc mRNA and protein was detected by qRT-PCR and Western blotting.Results The electrophoresis results of Eμ--VH-c-Myc-H2BmRFP gene fragment were in agreement with the predicted value of 4129 bp,and the electrophoresis of the vector DNA showed a band that was consistent with the pre-dicted value of 8561 bp.Enzyme digestion and DNA sequencing demonstrated that pEMIR was successfully constructed. Cells transiently transfected with pEMIR displayed the green and red fluorescence under inverted fluorescence microscope, and the expression of c-Myc was normal.Conclusion The lentiviral vector harboring c-Myc gene under the control of Eμ-VH promoter is successfully constructed,and the experiments in vitro demonstrate that it can be expressed in human normal cells and tumor cells.关键词
Eμ-VH启动子/c-Myc癌基因/慢病毒载体/弥漫大B细胞淋巴瘤Key words
Eμ-VH promoter/c-Myc oncogen/lentivirus vector/diffuse large B-cell lymphoma分类
医药卫生引用本文复制引用
陈琳,陈燕,肖东..Eμ-VH 启动子调控 c-Myc 表达慢病毒载体的构建及其体外功能[J].山东医药,2016,56(21):1-3,3.基金项目
广东省产业技术研究与开发专项资金资助项目（2013B060300013）。 （）
