Construction and characterization of a novel recombinant retroviral vector expressing mouse T-bet.
- Author:
Xuejie ZHANG
;
Jianhua ZHANG
;
Wei ZHANG
;
Jie GUO
;
Xuyu ZHOU
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line;
Genetic Vectors;
Interferon-gamma;
metabolism;
Mice;
Mice, Knockout;
Recombinant Proteins;
biosynthesis;
Retroviridae;
T-Box Domain Proteins;
biosynthesis;
T-Lymphocytes;
metabolism;
Transfection
- From:
Chinese Journal of Biotechnology
2014;30(10):1586-1593
- CountryChina
- Language:Chinese
-
Abstract:
In order to study T-bet function in mouse cells, a novel retroviral vector expressing mouse T-bet and reporter gene Thy1.1 was constructed. Retrovirus particles were then produced by transfection of the recombinant retroviral plasmid into a packaging cell line Platinum-E. The recombinant retrovirus played considerable infection ability. T-bet expression was then identified by FACS after infection of CD4+ primary T cells from T-bet knockout mouse with recombinant retrovirus. To determine if exogenous expressing T-bet has normal function, we checked the expression level of T-bet target gene, Ifng. IFN-y expression was upregulated in the T-bet knockout T cells infected with recombinant retrovirus. In conclusion, we successfully constructed an effective mouse T-bet recombinant retroviral vector.