Construction and expression of hIL-2/mGM-CSF fusion protein
- VernacularTitle:hIL-2/mGM-CSF融合蛋白的构建及表达
- Author:
Rong XIA
;
Xingmei LINLAI
;
Hongwei ZHOU
;
Al ET
- Publication Type:Journal Article
- Keywords:
hIL 2 mGM CSF Cytokine Fusion protein E.coli
- From:
Chinese Journal of Immunology
2001;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:hIL 2/mGM CSF fusion gene was constructed and expressed in E.coli.recombinant hIL 2/mGM CSF fusion protein had both biological activity of hIL 2 and mGM CSF.Methods:The hIL 2 and mGM CSF genes were amplified by PCR (splicing by overlap extension,SOE) and ligated with two prolines linkers.Then the fusion gene was cloned into vector pLY4 and PBV220.The recombinant plasmid were transfected into E.coli and expressed.Results:The sequence of hIL 2/mGM CSF fusion gene was correct the hIL 2/mGM CSF fusion protein was highly expressed in E.coli and that comprises 20% of total bacterial protein.The results of biological activity assay showed that the expressed product possessed both of the activities of hIL 2(4.5?10 5 U/mg) and mGM CSF(3.85?10 6 U/mg).Conclusion:Have successful constructed the recombinant hIL 2/mGM CSF fusion protein which possessed bioactivities similar to that of the natural hIL 2 and mGM CSF.
