Construction Of Plant Expression Vector With Human Endostatin Gene And Trasformation Of Tobacco

Endostatin, a carboxy-terminal 20KDa fragment of the non-collagenous (NC) 1 domain of collagen â…© â…§, is the first endogenous angiogenesis inhibitor entering human clinical therapy. It can inhibit endothelial cell migration and multiplication to reduce the vascularization of tumors and finally kill tumors. The great advantage of the agent lies in its less toxicity and not inducing drug resistance when used. The use of plant bioreactors for the production of recombinant pharmaceutical proteins is safe and inexpensive. Cultivating transgenic plants on an agricultural scale will produce almost unlimited supplies of recombinant proteins for uses in medicine. The plant expression vector pGE, carrying a human endostatin gene, was constructed. pGE was introduced into Agrobacterium tumefaciens by triparental mating. Tobacco leaf discs were transformed via Agrobacterium Mmefaciens mediated procedure. Transgenic tobacco plantlets grew in induction medium and plants resistant to kanamycin were obtained. PCR, Dot blotting and Southern hybridization analysis confirmed that human endostatin gene had integrated into the genome of tobacco.