Cloning And Expression Of E2Gene In The Process Of Wheat Infected By Puccinia Triticina

E2which is ubiquitin-conjugating enzyme in the plant ubiquitin-proteasome pathway plays animportant role. This study is carried out on the basis of the preliminary work. Yan et al. had establisheda SSH library of the reaction between the wheat and leaf rust, from which the TaE2EST was contained.The test on this basis, RT-PCR analysis and western blotting analysis were used to further investigaterelative expression of the TaE2in different time points after wheat inoculated leaf rust. And abioticstress and biotic stress function of this gene further verification. After that, the eukaryotic expressionvector was obtained and then transformed into the Arabidopsis thaliana. Then we obtainedoverexpression TaE2gene in Arabidopsis plants T1generation. The mainly results are as follows:1: The CDS of the TaE2gene was cloned by the method of RT-PCR from the wheat seedlings’leaves.2：Using bioinformatics analysis,we got the TaE2synthetic epitope peptide fragments and usingimmunity rabbits,we obtained higher specificity TaE2strong anti-rabbit antiserum.3: The varietal expression of TaE2in both mRNA and protein level at different hour’s post theinoculation of leaf rust was verified by RT-PCR and Western Blotting.4: Function of E2gene in different adversity was identified in accidence5: The eukaryotic expression vector pSuper1300+-35S::TaE2was obtained and then transformedinto the Arabidopsis thaliana. Then we obtained overexpression TaE2gene in Arabidopsisplants T1generation.All the results above made a foundation for the further study of the TaE2’s function and themechanism in the reaction between the wheat and leaf rust.