Constuction Of Plant Expression Vectors For Sense And Anti-sense Zinc Finger Proteins Gene Of Sugacane And Transformation Of These Genes Into Sugacane

Sugarcane(Saccharum officinarum L.) is one of the most important sugar crops, also it is an important chemical material for green energy source of ethanol, fibre, sugar chemical, feedstuff. In the long-term evolution, Plants generate a series of stress response mechanisms. Plant engineering strategies for abiotic stress tolerance rely on the expression of genes that are involved in signaling and regulatory pathways or genes that encode proteins conferring stress tolerance or enzymes present in pathways leading to the synthesis of functional and structural metabolites. The first gene type is regulator gene,while the two behind belong to effector gene.regulator gene can activate the expression of effector gene.Abiotic stress can activate the expression of some transcription factors(TFs) related to respond environment pressure, which can evoke the expression of a series of effector genes, so that's how the plants face the abiotic stress. Zinc finger protein belongs to an important transcription factor family that widely exists in both prokaryotic and eukaryotic organism. This kind of TFs participate in a lot of life activity, includes gene expression regulation, cell differentiation, embryo development, enhance the plants resistant ability to abiotic stress.According to the sequence of ShZP cloned from sugarcane by our laboratory,we designed special primers outside Open reading frame, introduced the cloned restriction site through PCR which respectively forwardly and reversely inserted between Prd29A promoter and NOS terminator of intermediate vector pCRBI, constructed the expression vector of trans-sense pCRBIShZPand anti-sense pCRBIantiShZP, the resistance marker genes of this plant expression vector is Bar gene.The two vector was transferred into Agrobacterium tumerfaciens EHA105 used to transform sugarcane embryonic callus by agrobacterium-mediated transformation method,after differentiation culture and continuous resistance screening,there were 43 trans-sense resistant plants and 41 antisense resistant plants,then got 10 trans-sense resistant plants and 8 antisense resistant plants through PCR testing of Bar gene.6 trans-sense resistant plants and 8-antisense resistant plants were tested through PCR testing between promoter region and target gene, we primarily prove that sense ShZP and anti-sense ShZP had been integrated into the sugarcane genome, all this Laid foundation for the latter functional analysis.