Preparation of human telomerase reverse transcriptase-immortalized MRC-5 cells
10.3760/cma.j.issn.1003-9279.2015.02.028
- VernacularTitle:人端粒酶逆转录酶永生化MRC-5细胞方法的建立
- Author:
Meng WU
1
;
Jianzhong YIN
;
Wuyang ZHU
;
Zhuozhuang LU
Author Information
1. 昆明医科大学公共卫生学院
- Keywords:
Cells,culcured;
Cytomegalovirus;
HIV-1 Revers transcriptase
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(2):177-179
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare immortalized MRC-5 cells by transduction of human telomerase reverse transcriptase gene (hTERT),which can be used for the routine propagation of human cytomegalovirus (HCMV).Methods The recombinant plasmid pLVX-EFla-hTERT was constructed by cloning hTERT gene into the vector pLVX-EFla-Tet3G,and hTERT gene was amplified from another plasmid pLVX-EFla-Tet3G.Lentvirus particles LVX-EF1a-hTERT were prepared by co-transfected 293T cells with pLVX-EF1 a-hTERT and Lenti-X HTX Packaging Mix.After that,Lentvirus particles were used to infect MRC-5 cells,then both the propagation and expression of hTERT in the passaged MRC-5T were examined.Results We obtained Lentvirus particles LVX-EF1a-hTERT and immortalized MRC-5 cells.Conclusion Method of human telomerase reverse transcriptase-immortalized MRC-5 cells will contribute to the construction of immortalized primary cells.
