Construction of lentiviral vector encoding CLEC4M and overexpression of CLEC4M in K-562 cells
10.3969/j.issn.1001-5256.2014.06.010
- VernacularTitle:CLEC4M慢病毒载体的构建及其在K-562细胞中的表达
- Author:
Yuanyuan WANG
;
Qinghe NIE
;
Ting ZHU
- Publication Type:Journal Article
- Keywords:
hepertitis C virus;
lentivirus infections;
dendritic cells
- From:
Journal of Clinical Hepatology
2014;30(6):518-521
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the lentiviral vector encoding CLEC4M and prepare K -562 cells with stable overexpression of CLEC4M.Methods The gene sequence of normal CLEC4Mwas cloned by reverse transcription PCR and then inserted into GV166 vector to construct GV166-CLEC4Mlentiviral expression vector,and then lentiviral packaging was performed by transfection of293T cells.The ob-tained lentiviral liquid was used to infect human leukemia cell line K-562.Real-time PCR and Western blot were used to detect the over-expression of CLEC4M in K-562 cells.Results Sequencing showed that the recombinant lentiviral expression plasmid GV166-CLEC4M was successfully constructed.Lentiviruses could efficiently infect K-562 cells,according to real-time PCR.CLEC4Mwas successfully o-ver-expressed in K-562 cells at mRNA and protein levels.Conclusion The construction of lentiviral vector encoding CLEC4M lays a foundation for further study of CLEC4M gene involved in HCV entry into host cells.
