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Expression， Purification of the MMP-9-PEX in E.coil and Its Effect on Invasive Capacity of Colonic Cancer Cells
Chun-Xia Yan, Jun-Lin He, Xue-Qing Liu, Yu- Bing Ding, Xue-Mei Chen, Ying-Xiong Wang*
Department of Heredity and Birth Health, The School of Public Health, Chongqing University of Medical Sciences, Chongqing, 400016, China
Abstract: Matrix metalloproteinase 9 hemopexin domain (PEX)， which locates the C-terminal， was amplified by RT-PCR and was inserted into expression vector pET32a. The recombinant plasmid was induced by IPTG for 4 h and a 42 kDa recombinant protein was produced. Amount of the fusion protein pET32a/PEX expression was 50% of total bacterial protein， mostly in form of inclusion， few in form of supernatant. Inclusion was dissolved in 8 mol/L urea and 10 mmol/L DTT， carried out affinity purification under denaturing condition. The expressed 42 kDa fusion protein is confirmed by SDS-PAGE. The pET32a/PEX expression vector was successfully constructed and highly expressed in E.coli. The purified pET32a/PEX protein can inhibitor the invasion of the colonic cancer cell in Transwell experiments and show a dose-dependent manner.
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