Prokaryotic Expression and Antifungal Activity of HumanαDefensin-5 Protein
10.3969/j.issn.0253-9896.2014.08.006
- VernacularTitle:人源性防御素HD-5的原核表达及抗真菌作用的初步研究
- Author:
Pingping ZHANG
;
Lirong YIN
;
Fang WANG
;
Bei SUN
;
Yan HUO
- Publication Type:Journal Article
- Keywords:
defensins;
recombinant fusion proteins;
Escherichia coli;
human alpha defensin 5;
prokaryotic expres-sion;
affinity purification;
antifungal activity
- From:
Tianjin Medical Journal
2014;(8):755-758
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.
