The Modification Of LEA Gene Vector Construction And The Transformation Of Upland Rice

Rice is the main food crops in the world. Dry rice, also known as upland rice, isanother kind of rice which is a cultivated rice type with a strong drought resistance andability to endure barren. Separation of drought resistance gene by biological technology,and through transgenic methods to cultivate new varieties of upland rice. This is aneffective way to increase crop yield. By using the method of agrobacterium-mediatedtransformation, this research has taken the ripe seeds of upland rice as the explants. Gettingthe reverse genetic-LEA gene transform into the plant genome, so as to obtain thetransgenic resistance of new upland rice varieties.In order to make the expression of LEA gene in the plants more efficient, we builtLEA gene into the pCAMBIA3301, than insert the ubiquitin promoter at the multiplecloning site, with using the bar gene as aselection marker gene. Than put the constructionexpression vector of pCAMBIA3301-LEA plasmid into the agrobacterium of Agl-1. Withusing the agrobacterium-transformation mediated, we can transfer the purpose fragmentsinto the upland rice. The achieved genetical modified (gm) upland rice will be identified atthe DNA level.To get the genetically modified (gm) seeds of upland rice for DNA purification, anddesign the primer of LEA gene, for PCR amplification. To preliminary screen the positiveplant which has already integrated the LEA gene into the upland rice genome. On the thirdgeneration, we can determine the insertion site of purpose gene through the Tail-PCRtechnology. And we can also appraisal the expression of purpose gene in the genomethrough the RT-PCR, to determine the stable genetic new strains of upland rice.