Expression and identification of recombinant fusion protein of thrombopoietin mimetic peptide and growth hormone in Pichia pastoris
- VernacularTitle:重组融合蛋白rTMP-GH在巴斯德毕赤酵母中表达的研究
- Author:
Song WANG
;
Junping WANG
;
Mo CHEN
;
Yang XU
;
Fang CHEN
;
Mingqiang SHEN
;
Aiping WANG
;
Yongping SU
;
Li KAI
- Publication Type:Journal Article
- Keywords:
thrombopoietin mimetic peptide;
growth hormone;
Pichia pastoris;
expression
- From:Journal of Third Military Medical University
2003;0(21):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To fulfill high-efficient expression of rTMP-GH,a recombinant fusion protein of thrombopoietin mimetic peptide and human growth hormone,in Pichia pastoris.Methods cDNA fragment coding for rTMP-GH was amplified by PCR,and subsequently cloned into the expression vector pPIC9K.The linearlized plasmid pPIC9K-rTMP-GH was transformed into Pichia pastoris GS115 cells and screened in MD/MM plates under pressure of G418.The recombinant fusion protein rTMP-GH was identified by Western blotting,and its biological activity was analyzed by its ability on colony formation of megakaryoblasts.Results The pPIC9K-rTMP-GH expression plasmid was successfully constructed.GS115 cells with high expression of rTMP-GH was screened out,and the rough purified rTMP-GH showed promotive effect on megakaryoblast colony formation.Conclusion High-efficient expression of rTMP-GH in Pichia pastoris is achieved.
