The Cloning And Functional Analysis Of Anion Channel TaVDAC1 And Chaperone TaTCP22

Wheat is one of the most important food crops in the world.The yield of wheat directly affects the survival of people,and the quality and yield of wheat are affected by many factors.Abiotic stress is one of the seriously factors that affect the yield of wheat.Therefore,it is very important to study the function of genes involved in signal pathway when suffered stress.Chaperone is a class of proteins that help other proteins to be folded,assembled,transported and degraded in the cell.At present,it was reported that TaTCP-1 protein part in various abiotic stresses,but the interaction protein with the TaTCP-1 is less.In order to provide a theoretical basis for the expression of molecular chaperone proteins involved in stress response signaling pathway,the TaTCP-1 was screened by using Gal4 system to screen the cDNA library.Using NCBI and Pfam to analysis the candidate clones,the discovery of most proteins involved in the signal transduction and energy metabolism processes.It contains chaperone protein gene TaTCP-22 and anion channel gene TaVDAC1.The major results are as following:1.Identification and analysis of Wheat TCP-1/cpn60 family.we identified 24 Wheat TCP-1/cpn60 genes divided into 2 groups.Promoter analysis indicated that TCP-1/cpn60 contained adversity stress.Based on RNA-seq data,TCP-1/cpn60 genes expressed in various tissue,it was also using RNA-seq data to examine the expression patterns of TCP-1/cpn60 under heat stress condition and drought stress.TaTCP57.3c showed higher expression levels in both heat and drought stress.2.Cloning and molecular characteristics of anion channel TaVDAC1 Gene.The phynogenetic analysis showed that TaVDAC1 belongs to Porin3 family,at low temperature,the expression level of TaVDAC1 was up-regulated,and was down-regulated by ABA,PEG,SA,and NaCl treatments.Subcellular localization demonstrated that TaVDAC1 locates mainly in cytoplasm,cytomembrane and nuclear.A 90 kDa of soluble fusion protein was successfully induced and identified as target protein by MS when pCold-TaVDAC1 was transferred into the E.coli strain BL2.TaVDAC1 transgenic plant germination experiment indicated that TaVDAC1 may involved in salt stress.3.Cloning and molecular characteristics of chaperone TaTCP-22.RT-PCR analysis indicate that TaTCP-22 was higher expressed in root.TaTCP-22 responses to ABA,PEG,SA,NaCl and low temperature.TaTCP-22 soluble fusion protein was successfully induced.In conclusion,the 38 candidate proteins may interaction with TaTCP-1,and most of the energy metabolism and plant signal transduction,contains a chaperone protein TaTCP-22 and an anion channel protein TaVDAC1.As follows the two genes were cloned and analysis the functions,it is helpful for the further function research of these genes.