Identification, purification, expression and cloning of fusion protein rhTum-5

AIM: To construct the recombinant expression vector of Tum-5 and to purify and identify the fusion protein rhTum-5. METHODS: A 264 bp of human Tum-5 gene fragment was synthesized and cloned into pQE-30 vector, an E.coli expression vector. The plasmid was transformed into E.coli DH5α and induced to express fusion protein rhTum-5 with IPTG. The expression of Tum-5 was detected by Tricine-SDS-PAGE and Western blot. RESULTS: A novel protein with expected molecular mass was expressed upon induction with IPTG. The expressed product accounted for about 40% of total bacterial proteins and showed good reactivity to anti-His tag antibody. CONCLUSION: Our successful cloning and expression of human Tum-5 gene and purification of rhTum-5 protein lay a basis for further study on the application of this protein to anti-angiogenesis in vitro and in vivo.