The Construction, Expression, Purification And Bioassay Of Neovasculature Targeted Recombinant Tum-5 Fusion Protein In E.coli

Angiogenesis is associated with several pathological disorders. Tumor growth and metastasis are dependent on angiogenesis. Components of vascular basement membrane are involved in regulating angiogenesis. tumstatin (the NC1 domain of α3 chain of type IV collagen) was identified as possessing anti-angiogenic activity, the anti-angiogenic activity of tumstatin was localized to the 54-132-amino acid Tum-5 domain, the activity mediated by αvβ3 integrin interaction in an RGD-independent manner, and the anti-angiogenic activity of Tum-5 is same as Tumstatin.Peptide NGR is a kind of tumor targeted peptide, which can home selectively to the vasculature of tumor, is selected from phage display library . To date, this high affinity characteristic of NGR-containing peptide to the has been widely employed to targeted deliver the radioelements, virus vector to the tumor neovasculature.Then we synthesis the sequence encoding Tum-5 residues and ligate NGR with C terminal of Tum-5 gene. We hope that the product of this fusion gene can maintain the anti-angiogenic activity effect of Tum-5 and furthermore homing Tum-5 to newly developed tumor vascular.In our research, we synthesized the gene encoding the amino acids 45-132 of tumstatin as tum-5 which includes the N-terminal 9 amino acids in addition to the previously identified anti-angiogenic domain (54-132 amino acids) . The...