Grape Resveratrol Synthase Gene Cloning And The Construction Of Highly Plasmid Expression Vector Of Sugar Beet

Resveratrol is one of a group of compounds called phytoalexins which mainly has the function of antibiosis, resisting oxidization, preventing platelet agglutination, protecting cardio-vascular, resisting cancer and so on. But the content of Res is low in plant under natural environment, so it is difficult and costly to use traditional method to get plentiful Res. (resveratrol synthase) RS is the final enzyme which plays a key role in the way of Res biosynthesis. In this paper, the RS gene was got by molecular biology method meantime the expression was reached in prokaryotic and the efficiently expressed genes was used to construct high expression of plasmid vector of beet In order to get high abundance of resveratrol At present the details has been made as follows:The genomic DNA was extracted from the selected six wild grapes in improving CTAB method. the resveratrol synthase gene CNRS1, CNRS2, CNRS4, CNRS5, CNRS6 which gene length 1179~1183bp were cloned by Suspension extension PCR method Has the outset codon and the termination codon, the integrity reading frame, coded 389~392 amino acids, With the known grape RS base nucleotide sequence homology respectively is 95% and 98%, 96% and 98%, 89% and 91.3%, 91% and 95%, 96% and 97.5%. Note that the cloned gene fragment is the integrated RS.All genes using molecular biological methods were cloned to prokaryotic expression vector pET-30a-CLRS and transformed into Escherichia coli BL21 (DE3) and the resistance selected IPTG induced expression and SDS-PAGE electrophoresis the protein were obtained with the gene encoding and the size is equal to the theoretical, Protein expression production increased gradually with the induction of time The highest abundance in 4h in which the highest relative expression is CNRS2 gene.With the efficient expression of CNRS2 gene constructed beet plasmid high expression vector pSARCNRS2 Using molecular biological methods, For further research on breeding resistant varieties of sugar and as a bioreactor expressed to resveratrol laying the foundation.