High efficient expression of human endostatin in E.coli and its antiangiogensis activity |
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| Authors: | ZHANG Ge KE Xi-song DING Ai-ling YING Wei YANG kun ZHU Zhen-yu |
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| Institution: | 1. The Laboratory of Molecular Medicine of Sun Yat-sen University of Medical Sciences, Guangzhou 510089, China;
2. Department of Biochemistry, Chongqing University of Medical Sciences, Chongqing 400.016, China;
3. Department of Biochemistry, Yunyang Medical Collage, Shiyan 442000, China |
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| Abstract: | AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors. |
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| Keywords: | Endostatin Antibodies Biological activity |
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