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Large subunit of human heparanase protein in E. coli fusion expression
PAGE \* MERGEFORMAT 15
Large subunit of human heparanase protein in E. coli fusion expression
Authors: Li Yanjie, LI Ji-Xue, Ling Yan, Tang state, Lin Yan, Zhu Hou Chu
[Keywords:] human liver acetyl
Cloning, expression and characterization of human heparanse Cterminal subunit protein in E. coli
[Abstract] AIM: To construct the expression vector for heparanase (HPA) Cterminal subunit and obtain enough protein for further study. METHODS: Full encoding gene of heparanase protein was isolated from the human platenta cDNA library and its expressing vector pGEX2TK50 ku HPA was constructed by cloning its Cterminal subunit gene into the pGEX2TK vector. After transformed by the expression vector, E.coli BL21 (PlyS, DE3) was induced to express the target protein. RESULTS: E.coli BL21 (PlyS, DE3, pGEX2TK50 ku HPA) began to express the target protein fused by GST protein and Cterminal subunit protein after 1 h induction and kept producing it until 3 h after induction when the target protein reached a maximum. No degradation of produced protein was observed in the hetero E.coli cells. The target protein was expressed mostly in inclusive bodies. When growing at lower temperature, some fused protein was detected soluble in cell lysis. CONCLUSION: The Cterminal subunit protein of heparanase fused with GST protein has been successfully expressed in E.coli BL21 (PlyS, DE3). The protein may be a good antigen in preparing the antibody for heparanase and in further study of its function.
[Keywords] heparanase; Cterminal subunit; pGEX2TK; gene expression
[Abstract] Objective: Heparanase in tumor metastasis, angiogenesis, histogenesis play an important role in such processes, its carboxy-terminal large subunit protein is an important component of the enzyme, but because of this protein in the body content of less easily biodegradable it is difficult to directly from within the organization to be a certain amount of protein. In this study the use of engineering bacteria to ex
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