Functional Analysis And Transgenic Research Of Four Cold-and Drought-induced Genes From Ammopiptanthus Mongolicus

We obtained four cold- and drought-induced genes:AmRRM, AmZFPG, AmSTZF, AmCaM from Ammopiptanthus mongolicus and researched their function. We analysed cDNA sequence,amino acid properties, protein conservation, phylogenetics, protein structure and function and forecasted that these genes are related to stress response.We also analysed gene expression of Ammopiptanthus mongolicus leaves incubated at 4℃and under drought stress by RT-PCR. RT-PCR analysis shows that the expression of genes is related to response to drought and chilling stress. In order to further study gene function, we constructed these four genes to vector, transformed them into E. coli BL21 to gain target protein.DNA sequencing showed that these genes were transformed into E.coli successfully and SDS-PAGE results indicated that AmRRM and AmCaM can express in E.coli. And then, we prepared antibody with purified AmRRM protein and analysed the expression of AmRRM in plant by western blotting.Simultaneously, we constructed AmRRM,AmZFPG,AmSTZF to 2300 vector, transformed them into agrobacterium LBA4404 and infected the leaves of wild type tobacco. We incubated wild type and transgenic tobacco at 4℃and measured the electrical conductivity.The results showed that the cold resistance of the transgenic tobaccos was higher than the wild type tobacco. So we concluded that AmRRM,AmZFPG amd AmSTZF genes which obtained from Ammopiptanthus mongolicus might play roles in protecting plant cells from cold stress injuries.