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Vascular endothelial growth factor (VEGF) as a powerful blood vessel growth medium , not only specifically acts on vascular endothelial cells , stimulate cell division and proliferation and migration , and plays an important role in vascular maturation and remodeling , promote therapeutic angiogenesis . VEGF-C is a specific blood vessels , lymphatic endothelial cell regulatory factors , the main role is to increase the permeability of vascular endothelial cells , mediated lymphatic endothelial cell chemotaxis , cell proliferation , and lymphatic hyperplasia . Many studies suggest , is closely related to tumor lymphatic metastasis and VEGF-C . Thus , VEGF-C tumor lymphatic metastasis treatment to provide a new way . In this study, by constructing the VEGF-C gene pGEX / VEGF - C recombinant , expressed in Escherichia coli , to obtain high-level expression of recombinant VEGF-C protein to further study its structure and function , and in-depth understanding of tumor pathogenesis , promote anti-tumor the development of the drug market . The main contents and results are as follows : 1 , the test chemical synthesis VEGF-C gene , inserted into the pGEX-6p-1 fusion protein expression vector to construct the recombinant plasmid pGEX / VEGF -C . Transformed into E. coli E. coli BL21 transformants screening of high- efficiency , shake flask cultures efficient expression of the protein was induced with IPTG . The expression product was purified using glutathione - agarose affinity chromatography and the protein content was measured by Lowry's method . Successfully constructed prokaryotic expression plasmid pGEX / VEGF -C ; SDS - PAGE analysis of induced by IPTG , the recombinant expression plasmid pGEX / VEGF-C expression in E. coli fusion protein with a molecular weight of about 61kD ; through the optimization of the induction conditions protein expression reached a peak after 16 hours in 16 ℃, 0.4mM IPTG induction . 3, a recombinant fusion protein expressed in E. coli after glutathione - agarose affinity chromatography purification to obtain a higher concentration of VEGF-C-GST fusion protein, while digestion with a high purity of the VEGF-C protein , and the protein content of 20 mg / ml. Lay the foundation for the training of the crystal , the preparation of monoclonal antibodies , to study the protein structure and function .
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