Genetic Transformation Of Plant And Yeast With RScDNA Cloning From Peanut

Res (resveratrol) is a kind of plant endotoxin. It is a kind of composion with health care function existing on several dozen food and one of medicine. Res mainly has the function of resisting cancer, protecting cardiovascular, regulatin estrogen, resisting hyperplasia,leading cell to dying, antibiotic and so on. Res is low in plant, so it is difficulty and costly to only use traditionally withdraw method to get plentiful Res. Research shows that RS (resveratrol synthase) is the only necessary enzyme and the last one enzyme in the way of Res synthesizing. In this paper, we use the method of molecular biology to get RScDNA and genetic transformation plant and Pichia.pasteoris separately. At present, we have already made the following progress.RScDNA was cloned according to over-hang extension PCR protocol with peanut genomic DNA as template. Through sequence analysis showed that RScDNA was correctly obtained. One hand, contrast RScDNA sequence with AB027606, the homology is up to 95%.On the other hand , amino acid contrast to p20178, the homology is more than 98%.RScDNA gene confirmed correctly was integrated into a plant express vector, PBI121, from which GUS gene sequence had been removed .The vector ,PBI121-L containing RScDNA under control of CaMV35s promoter and NOS terminator was constructed and introduced into LBA4404. Peanut and carrot hypocotyl were infected with Agrobacterium.At the same time, RScDNA gene was cloned into pPIC3.5K named pPIC3.5KL. The restriction analysis results showed that the RScDNA gene was ligated correctly into pPIC3.5K. With the electricity dash the recombinant plasmid was switch to the Pichia.pasteoris strain GS115. The strain with recombinant plasmid was chosen by PCR.