Construction Of Dehydrin-BDN1 Binary Expression Vector And Agrobacterium Mediated Leaf-disk Tobacco Transformation

The studies of drought-tolerant plants have turned from the plant physiology to molecular biology with the development of plant genetic engineering. The methods of molecular biology about drought-tolerant plants are mainly genetic engineerimg technology including the clone of drought-tolerant genes, the identification of drought-tolerant genes and getting transgenic drought-tolerant plants. Dehydrins have been the most commonly observed, among proteins that accumulate in plants in response to dehydrative forces or low temperature. Dehydrins have become the highlight in the field of plant tolerance to drought. In this experiment, the gene桰IDN1 was cloned from the resurrection plants. Then BDN1 was used to construct the binary expression vector pBI121BDN1.The BDN1 is promoted by the promoter--CaM35S. And the monocotyledon expression vector梡AHC25BDN1, the objective gene-BDN1 is promoted by the Ubiquitin--promoter which was cloned from the maize. We use the gene Trehalose-6-phosphate synthase (TPS) as the expression gene to construet the monocotyledon expression vector梡AHC25TPS, TPS is promoted by the Ubiquitin-promoter. The binary vector plasmids-pBI121BDN1 were transformed into tobacco by the Agrobacterium mediated leaf-disk methods. After the transformation, 8 plants among 20 plants which can grow up in the Kan selective mediums identified by PCR have the 800pb objective gene-BDN1 DNA band, that is, the result is positive. Then the 8 plants were analyzed by DIG Southern-blotting, 2 plants had corresponded DNA blotting band.