Construction and function of a lentiviral vector harboring c-Myc gene under the control of Eμ-VH promoter
CHEN Lin
CHEN Yan
XIAO Dong
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.
Keywords：Eμ-VH promoterc-Myc oncogenlentivirus vectordiffuse large B-cell lymphoma
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 1-3 )