Expression and functional study of human recombinant chemokine-like factor I in Drosophila S2 cells
- VernacularTitle:人重组趋化素样因子1在果蝇S2细胞中的表达和功能研究
- Author:
Yingmei ZHANG
;
Ting LI
;
Yaxin LOU
;
Wenling HAN
;
Dalong MA
- Publication Type:Journal Article
- Keywords:
Chemokine-like factor 1;
Chemokine;
C2C12 cells
- From:
Chinese Journal of Microbiology and Immunology
2008;28(6):481-485
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express human chemokine-like factor 1 (CKLF1) in Drosophila S2 cells and study its function. Methods The pMT/V5-His-CKLF1 expression plasmid was constructed and transfected into Drosophila S2 cells. The positive clones were selected through PCR and RT-PCR. The culture medium was analyzed by Western blot with anti-CKLF1 polyclonal antibody. Chemotaxis and MTT assays on human peripheral blood and C2C12 cells, respectively, were then carried out with the medium. Results CKLF1 was transcribed efficiently in S2 cells. The expressed CKLF1 protein could be detected in the culture supernatant by Western blot, which showed weak chemotactic activity on both human peripheral blood neutrophils and lymphocytes as well as enhancing effect on the proliferation of C2C12 cells. Conclusion CKLF1 was expressed successfully in Drosophila S2 cells and secreted into the culture medium. The recombinant CKLF1 expressed in Drosophila cells can chemoattract leucocytes and promote the proliferation of C2C12 cells.
