Studies On Cloning、Expression Of UFGT Genes And Induced By UV-C Irradiation In Grapes

Anthocyanins, not only can improve the ability to adapt and resist stress, but also is good for health, is one of the important factors in determining the quality and sensory of wine. UFGT is the last enzyme of anthocyanins’ synthesis in plants, affects the content of anthocyanins. In this study, a full length ufgt cDNA was cloned from grape berries (Vitis vinifera L.cv. Cabernet Sauvignon),then prepared polyclonal antibody. The grapes were treated with UV-C irradiation regularly at the development stages, and studied the influence of ufgt expression pattern on the level of transcriptional and translational to the accumulation of anthocyanins.The main results are as follows:A full length ufgt cDNA with 1371 bp was cloned from grape berries, then build the cloning vector. The cloning vector was introduced into the pET-30a expression vectors by restriction enzyme digestion. UFGT protein was highly expressed and purified through optimizing protein expression. Grape UFGT antibody was producted by immunizing a New Zealand rabbit with the purified UFGT protein as an antigen.Grape berries{Vitis vinifera L. cv. Cabernet Sauvignon) were used as materials, studied the influence of UFGT to the content of anthocyanins under CK and UV-C 10 min. Using the Real-Time PCR to detect the law of ufgt gene expression during grape development, UV-C 10 min does not induce the expression of ufgt gene, and the law is similar with CK. Using the western-blot to study the content of UFGT protein during grape development, UV-C 10 min does not change the laws of the accumulation of UFGT protein, but significantly induces the accumulation of UFGT protein. Using the HPLC and the spectrophotometer method to detect the activity of UFGT enzyme and the content of anthocyanins during grape development, the activity of UFGT and the content of anthocyanin are higher under UV-C 10 min. Analysis the correlation of ufgt gene expression, the content of UFGT protein, the activity of UFGT and the content of anthocyanins.Analysis the information of UFGT bioinformatics, we know that UFGT protein is composed of 456 amino acid residues, the molecular weight is about 50 kDa, may contain signal peptide and transmembrane domain prediction, the secondary structure is constitute by 35.53% α-helix,16.67% sheets extended strand and 47.81% random coil. Analysis the affinity, the UFGT protein of Vitis vinifera L. cv Cabernet Sauvignon homology is the highest with grape(Vitis amurensis), then is strawberry, and the homology with cornis is the lowest.