Preliminary Studies On Transformation Of Sj23 Gene Into Medicago Sativa

Schistosomiasis is a worldwide parasite communicable disease,which endangers the human and livestock health.The development of schistosomiasis vaccine has a great economic value and social significance. Transgenic plant-derived vaccine developed in recent years,and ompare with other vaccines,it's cheaper,safer and more efficacious,and become important in vaccin research. Medicago sativa is a kind of perennial leguminous caudex and is the best and most important leguminous in the world.In order to develop transgenic plants Schistosomiasis vaccine,we transformed the Sj23 gene into a Medicago sativa throuth Agrobacterium-mediated,and identificated the transgenic callus by PCR and Western blot.The main results are as following:1. Amplified the Sj23 by PCR and TA clone,then construct the recombinant prokaryotic plasmid pCAMBIA1303-Sj23.2. Transformed the pCAMBIA1303-Sj23 into Medicago sativa through Agrobacterium- mediated.Induced the callus with the hormone of 2 mg/L 2,4-D and 0.5 mg/LKT. Access to 96% of the callus induction rate.By the method of delay screening,we got 120 resistant callus with the optimum selection pressure of 50 mg/L hygromycin.3. Extracted the hygromycin-resistant callus'DNA and PCR with the specific primers of Sj23,we got 68 positive callus.Chose 10 positive callus on random and extracted theri protein,by SDS-PAGE we found 2 of them expressed a 23kDa protein which no exist in the negative callus.By Western blot,we found this protein could be recognized by the serum from mice infected with Schistosoma japonicum,which means the Sj23 may expressed in the Medicago sativa callus.