Over-expression vector construction of human DcR3 gene and its validation
10.3969/j.issn.1000-484X.2016.10.018
- VernacularTitle:人DcR3表达载体的构建及验证
- Author:
Liulan PAN
;
Shengnan JIA
;
Jingting MA
;
Jinghua TAI
;
Zhenjing JIN
- Publication Type:Journal Article
- Keywords:
DcR3;
LX-2;
Eukaryotic expression;
Clone
- From:
Chinese Journal of Immunology
2016;32(10):1491-1495
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct the human DcR3 expression vector and verify its expression in vitro. Methods: 915 bp human DcR3 gene CDS was amplified from porcine lung tissues,and was cloned into eukaryotic expression vector pEF1a-IRES-DsRed-Express2 which show red fluorescence. And then pEF1a-IRES-DsRed-Express2-DcR3 was transfected into LX-2 cells by FuGene HD. Expression of mRNA and protein lever of Human DcR3 were detected by RT-PCR and Western blot. Results:The levels of DcR3 gene transcription and translation in the hepatic stellate cells were significantly increased after transfection with pEF1a-IRES-DsRed-Ex-press2-DcR3 by RT-PCR and Western blot analysis. Conclusion: DcR3 expression vector was successfully constructed and highly expressed in LX-2 cells.
