Cloning, Bioinformatics Analysis And Prokaryotic Expression Of SAMS Genes In Vitis Vinifera

In the process of growth and development, plant often encounters biotic and abioticstresses such as pests, disease, drought, injury, low temperature and so forth. Those adversitystresses will affect the growth and development of plant, leading to decrease of productionand quality. In currently, with the increas of population and degradation of environment, as animportant fruit in the world, grape is facing the stress problems in the cultivation. Therefore,cultivating new variety of grape which will resist or tolerant to the stresses has become one ofthe best ways to solve the problem.In this study, S-adenosylmethionine synthetase (SAMS) genes were cloned from V.vinifera. The squences of cloned genes were analyzed and then were expressed in prokaryoticE.coli. The results are as follows:1. Retrievaing and analyzing genome database of grape, we found five SAMS genes withentire open reading frame (ORF). Then results of preliminary bioinformatics analysis showedthat all of the five SAMS genes contain the specific N-terminal domain, middle domain andC-terminal domain of the gene. The proteins are not Secreted proteins and Membrane proteins.The secondary structures were dominated by the α-helix and random coil. We found that theSAMS gene of grape had a very high homology with the gene of Arabidopsis’. Those resultshave important theoretical significances for the further study on the structure and function ofVvSAMS gene.2. Two members of SAMS family with entire open reading frame (ORF), VvSAMS1andVvSAMS3, were cloned from leaf of Vitis vinifera.3. After the grape were treated under different stresses (SA, MeJA and mechanical injury), weanalyzed the changes of relative expression levels of VvSAMSs, by Real-time PCR method.The results showed that, under the stresses, the expression levels of VvSAMSs showed someobvious changes. The results implied that the expression of the VvSAMSs was responded tothe stresses.4. The VvSAMS genes were constructed into an expression vector and then transformed inE.coli BL21. The results of SDS-PAGE showed that recombinant protein expressedheterologously in the E.coli after induction.Cloning, analysis and prokaryotic expression of VvSAMS Genes built a foundation forthe further research on the involvement of stress resistance mechanisms and function of thegenes.