Construction of prokaryotic expression plasmid of fusion protein including porin A and porin B of Neisseria gonorrhoeae and its expression in E. coli.
- Author:
Fang LIAO
1
;
Qifa SONG
;
Mufen WAN
Author Information
1. Department of Microbiology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Humans;
Neisseria gonorrhoeae;
genetics;
Plasmids;
biosynthesis;
genetics;
Porins;
biosynthesis;
genetics;
Prokaryotic Cells;
metabolism;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Transfection;
Vaccines, Synthetic
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2004;24(5):417-420
- CountryChina
- Language:English
-
Abstract:
In order to provide a rational research basis for clinical detection and genetic engineering vaccine, plasmid pET-28a (+) encoding both Porin gene PIA and PIB of Neisseria gonorrhoeae was constructed and a fusion protein in E. coli DE3 expressed. The fragments of PIA and PIB gene of Neisseria gonorrhoeae were amplified and cloned into prokaryotic expression plasmid pET-28a (+) with double restriction endonuclease cut to construct recombinant pET-PIB-PIA. The recombinant was verified with restriction endonuclease and sequenced and transformed into E. coli DE3 to express the fusion protein PIB-PIA after induced with IPTG. The results showed PIA-PIB fusion DNA fragment was proved correct through sequencing. A 67 kD (1 kD= 0.992 1 ku) fusion protein had been detected by SDS-PAGE. It was concluded that the fusion protein was successively expressed.
