Cloning And Function Analysis Of The Promoters Of Drought-induced Genes From Maize

Drought ecological crisis has seriously affected the human sustainable development, food crops and vegetation can grow under drought tolerant and water stress environment is necessary in agricultural development. Higher plant gene expression regulation has become the hotspot of molecular biology research field and in the preface, while the promoter is an important element in regulating gene expression. Study the structure and function of promoters depth, establish empty and measured 3d regulation system used for plants genetically modified expression, can provide new ideas for functional genomics research and accurately adjusting plant metabolism through genetic engineering technology. Because constitutive promoters are not effectively controlled to express gene and are not controlled purpose gene expression from time and space. So study and utilization the induced and organization-specific promoters are particularly important. Maize is chinese important food crops, while drought stress is one of the important factors to influence maize yield. However, the research about maize drought induced promoters is less at present. This experiment obtain conclusions through maize drought induced gene ZmCIPK1, ZmLOC1 promoters.1. Search for drought induced genes ZmCIPK1, ZmLOC1 in NCBI website and analysis the 5 'section of the two genes by bioinformatics method.Finding promoter sequences have the typical TATA - box and CAAT-box, according with the eukaryotic promoters basic features. Also found the cis-regulatory elements of drought induced, such as MYB recognition sites, MYC recognition sites, and some related with salt induced and cold induced recognition sites. Forecasting the two promoters have the characteristics of drought induction.2. Construct plants expression vector pCAM∷PZmCIPK1,pCAM∷PZmLOC 1, transforme corn young embryo and tobacco callus through the method of agrobacterium-mediated . To analyze the instantaneous of GUS gene expression using the method of GUS histochemical staining. Find promoter PZmCIPK1active increase markedly after induction by drought, while promoter PZmLOC1 is not obvious.3. Obtain transgenic tobaccos by blades reforming process, and analyze the stability of GUS gene expression with the method GUS histochemical staining. Results show that promoter PZmCIPK1, PZmLOC1 can stable expresse in transgenic tobaccos. Transgenic tobacco activity increase apparently through induction of dry.4. In order to research the express way and the main control organization regional of promoters PZmCIPK1, PZmLOC1 and to analysis promoters by lossing 5 'section. Constructe plants expression vector respectively, pCAM∷PZmCIPK2,pCAM∷PZmCIPK3,pCAM∷PZmCIPK4,pCAM∷PZmCIPK5 and pCAM∷PZmLOC2,pCAM∷PZmLOC3,pCAM∷PZmLOC4,pCAM∷PZmLOC5. Fluorescence quantitative detection shows that the strongest activity section of promoter PZmCIPK is PZmCIPK5, while the strongest active section of promoter PZmLOC1 is PZmLOC5.