Preparation and bioactivity assay of mIL-4-SA fusion protein / 中国生化药物杂志
Chinese Journal of Biochemical Pharmaceutics
;
(6): 90-93, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-402723
ABSTRACT
Purpose To prepare streptavidin-tagged mouse interleukin-4(mIL-4-SA)bifunctional fusion protein and to study on its bioactivity.Methods The mIL-4 gene was cloned by RT-PCR and cloned into pET21 vector to get mIL-4-SA-pET21 expression plasmid.The mIL-4-SA fusion protein was expressed in BL21 (DE3)host bacteria and purified through the Ni-NTA affinity chromatography and refolded by dilution and dialysis.The effect of mIL-4-SA fusion protein on mouse thymocytes proliferation was evaluated by MTY.Flow cytometric analysis was performed to detect the mIL-4-SA fusion protein on the biotinylated B16F10 tumor cells.Results The mIL-4-SA-pET21 vector was successful by constructed and the mIL-4-SA fusion protein was expressed in BL21(DE3)at about 35%of total bacterial proteins.The purity of mIL-4-SA Was about 95% through Ni-NTA.The mIL-4-SA fusion protein exhibited bifunctional activities,i.e.,stimulative effect for mouse thymocyte proliferation and SA-mediated high-affinity binding to biotinylated cell surfaces(anchoring modified rate Was about 96.69%).Conclusion The mIL-4-SA fusion protein was expected to be developed for the treatment of tumors.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Biochemical Pharmaceutics
Year:
2010
Type:
Article
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