ABSTRACT
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. 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. 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 micri g/L VEGFR-1-II protein was produced. The function of this protein was confirmed by inhibition of HUVEC cells growth. Since this protein inhibited the angiogenesis in vitro it may be consider as an efficient anti-angiogenesis factor