cloning and expression of vascular endothelial growth factor receptor-1 in escherichia coli and analysis of its growth inhibition effect on human umbilical vein endothelial cells

objective: angiogenesis a process that results in neo-vascularization is an essential stage in growth of solid tumors and the formation of metastases. vascular endothelial growth factor (vegf) and its receptors, vegfr1 (flt-1) and vegfr2 (kdr/flk-1), are the major regulators for tumor angiogenesis. recent studies showed that second domain of vegfr-1is a key factor for vegf/vegfr-1 interaction. materials and methods: in this study, after rna purification and cdna synthesis, the second domain of vegfr-1 (vegfr-1-ii) was amplified by pcr and cloned in t/a cloning vector. in order to increase the expression of the protein, we sub-cloned the gene into pet22b(+) and transformed the construct in rosseta-gami 2, an efficient host for expression. the expression was induced by iptg and confirmed by sds-page and western blotting. the recombinant protein was purified by imac column and the growth inhibition of human umbilical vein endothelial cells (huvec) was analyzed by the recombinant protein. result: the results of sds-page and blotting confirmed the protein purification accuracy. the recombinant protein concentration was determined by bradford protocol. the results showed that nearly 300 µg/l vegfr-1-ii protein was produced. the function of this protein was confirmed by inhibition of huvec cells growth. conclusion: since this protein inhibited the angiogenesis in vitro it may be consider as an efficient anti-angiogenesis factor.