Secretory Expression of the Fusion Protein IFN?-HSA in Pichia pastoris
- VernacularTitle:人?干扰素-血清白蛋白融合蛋白在毕赤酵母中的分泌表达
- Author:
Jian-Yong LEI
;
Lian-Feng ZHANG
;
Jian-Liang YANG
;
Jian JIN
;
- Publication Type:Journal Article
- Keywords:
Interferon?Pichia pastoris Fusion protein
- From:
China Biotechnology
2006;0(07):-
- CountryChina
- Language:Chinese
-
Abstract:
Overlapping PCR technology was employed to splice IFN?and HSA genes in vitro. The spliced gene was inserted into Pichia pastoris secretory vector pPIC9K. The IFN?-HSA gene was designed for secretory expression under the control of promoter AOX1 and Mat a signal peptide in pPIC9K. The recombinant plasmid pPIC9K/IFN?-HSA was linearized by restriction enzyme SalI and transformed into Pichia pastoris KM71 by electroporation. The recombinant strains identified by G418 selection and confirmed by PCR analysis were induced by methanol to express fusion protein IFNp-HSA. SDS-PAGE and Western blot analysis of the fusion protein showed that the expressed fusion protein IFNp-HSA with an apparent 90kDa molecular weight had the antigenicity of HSA. The specific activity of culture supernatant was about 640IU/ml assayed by the standard amiviral activity test on WISH cells challenged with VSV virus.
