Overexpression of Mature Human Tissue Kallikrein Fusion Protein in E. coli

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Chinese Journal of Biochemistry and Molecular Biology ›› 2003, Vol. 19 ›› Issue (03) : 312-316.

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Overexpression of Mature Human Tissue Kallikrein Fusion Protein in E. coli

LI Ti yuan, DU Gong , CAI Xiao yan, SHI Zhi lin, HUANG Rui fang, CHEN De heng, DAI Yong, XIAO De ming

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Human tissue kallikrein gene fragment encoding mature kallikrein was amplified and cloned into pET 28(b) and pET 20(b) plasmid with C terminal 6×His Tag. The recombinant fusion protein was overexpressed in BL21(DE3), BL21(DE3)plysS and HMS174(DE3) under the induction of IPTG. SDS PAGE showed that the kallikrein fusion protein were produced at a level of approximately 26% and 10% of the total cellular protein in E. coli BL21(DE3) plysS by pET 28(b) and pET 20(b), respectively. The relative molecular weight of the expressed protein was 30 kD. Western blot analysis indicated that the expressed kallikrein had the antigenicity to human serum kallikrein. The kallikrein fusion protein in the un purified solution showed low activity to hydrolysis BAEE by using potentiometric method. The overexpressed human tissue mature kallikrein could be used for the development of genetic engineering products and further study of its biological property.

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kallikrein / overexpression / E. coli

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LI Ti yuan, DU Gong , CAI Xiao yan, SHI Zhi lin, HUANG Rui fang, CHEN De heng, DAI Yong, XIAO De ming. Overexpression of Mature Human Tissue Kallikrein Fusion Protein in E. coli[J]. Chinese Journal of Biochemistry and Molecular Biology, 2003, 19(03): 312-316

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Received	Revised	Published
2003-06-20	2003-06-20	2003-06-20
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2003-06-20

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