Study on transforming the gene encoding chimera SBR-CT^DA1 into tomato

Objectives: The propose of the study was to transform the plant expression vector pROSB carrying the gene encoding chimera SBR-CT^DA1 and bar gene into tomatoes, and build high efficient regeneration and transformation system of tomatoes. Methods: (1) The plant expression plasmid pROSB containing the gene fragment encoding chimera SBR-CT^DA1 and bar gene were constructed using genetic engineer technology. (2) Plasmid pROSB were mobilized into Agrobacterium tumefaciens LBA4404 by the freeze-thaw method to obtain Agrobacterium binary vector AT-RSB(pROSB,pLAB4404).(3)The condition of transformation including the concentration of ZT,IAA and antibiotic were ascertain by observing the differentiation and regeneration of tomato explants in grads concentration. (4)Tomatoes were transformed by leaf-disc cocultivation method using AT-RSB. (5) Positive transgenic plants were selected by PCR. Results: (1) The plant expression plasmid pROSB carrying the gene encoding chimera SBR-CT ^DA1 and bar gene was constructed successfully. (2)Agrobacterium binary vector AT-RSB(pROSB,pLAB4404) was constructed. Plasmid pROSB was reproduced stability in Agrobacterium tumefaciens AT-RSB.(3)ZT(2.0mg/L) and IAA(0.2mg/L)had great effect on the formation of callus and shoots; the effective concentration of kanamycin was 25-75mg/L;the appropriate concentration of Cefn(Cefotaximum Natricum and Sulbactamum Natricum) was 300mg/L.(4) Transgenic seedlings were obtained under the press of kanamycin and glufosinate(PPT). The 1.9kb target fragment was amplified from transgenic plant DNA. The gene fragment encoding chimera SBR-CT^DA1 and bar gene were transformed into tomatoes. Conclusion: It is feasible to transform the gene of causing caries of streptococci mutants to tomato, which may provide useful experiment foundation for edible plant vaccines.