Study On Agrobacterium Tumefaciens-Mediated Genetic Transformation Of Sweetpotato (Ipomoea Batatas(L) Lam.) With ANTI-CTLA-4-HS83 Gene

Plant-based expression of pharmaceutical proteins is a cost-effective alternative to traditional large-scale production systems such as microbial fermentation,insect and mammalian cell cultures,and transgenic animals.Plants are the most economical makers of biomass using only sunlight,water,carbon dioxide and minerals.They offer a proper modification system for eukaryotic proteins,such as folding,post-translational modification,and sub-cellular targeting;moreover,they avoid animal cell-culture contaminants such as mammalian pathogens and extraneous viral or bacterial components.Since the human growth hormone was expressed in transgenic tobacco in 1986,many more pharmaceutically relevant proteins have been successfully produced in plants.The proteins expressed in plants can be extracted from whole plant parts or organelles and used as a source for medicinal applications.Production of sweetpotato(Ipomoea batatas(L)Lam.)ranks fifth after that of rice,wheat,maize and white potato,among the food crops of the world.Sweet potato is one of the most important crops for developing countries,where it ranks second according to its production value,and fifth according to its calorie contribution for humans.Sweet potato have a high production yield of biomass,need less nutrients from the soil than most other crops.Establishing sweet potato as a production site for the delivery of recombinant proteins is worth striving for.Single chain fragment variables(scFv)anti-CTLA-hs83 is one kind of target fragments for immunotoxins,it can selectively link the activated T cells, possesses the features of specificity,high activity and low immunogenicity.So it maybe have good prospect for prevention and cure of transplant rejection. In this study,we describe an agrobacterium binary vector carrying single chain variable fragment(scFv)anti-CTLA- hs83 gene was successfully constructed based on pCAMBIA1301,then transformed sweet potato plants with this binary vector using A.tumefaciens-mediated transformation,30 hygromycin resistant plants were obtained.Hygromycin resistant plants were tested by polymerase chain reaction(P.C.R.),the ruslts showed that 17 of them may be tansgenic plants.