华中科技大学学报(医学英德文版)
華中科技大學學報(醫學英德文版)
화중과기대학학보(의학영덕문판)
JOURNAL OF TONGJI MEDICAL UNIVERSITY
2004年
5期
417-420
,共4页
廖芳%宋启发%万沐芬
廖芳%宋啟髮%萬沐芬
료방%송계발%만목분
Neisseria gonorrhoeae%porin B%porin A%prokaryotic expression plasmid recombinant%fusion protein
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.
