Cloning And Functional Analysis Of PIP5K Gene Promoter In Zea Mays

Promoters play an important role in plant gene expression. Provide a theoretical basis for the cloning and functional study of plant gene promoter contribute to the understanding of signal transduction pathways and gene expression regulation mode, for plant genetic engineering research. Tissue-specific promoter as a promoter to start the expression of genes in a particular organization and a certain developmental stage, can reduce unnecessary waste. PIP5K gene family as the root of a specific transcription factor, plays a vital role in coordinating the growth of plants, in particular, to respond to environmental factors, the initiation and promotion of root hair tip growth functions.In this study, the PIP5K gene sequence by bioinformatics analysis, and cloned the PIP5K gene promoter, and promoter deletion experiments, the genetic transformation of rice and rice tissue GUS staining experiments, the results of the following:First, according to a known reporter gene analysis and forecasting, to design specific primers, maize B73genomic DNA as template, amplified the part of the gene promoter, named P2036. P2036promoter sequence on the promoter cis-acting elements forecast website analysis, predicted the existence of the TATA box, CAAT box, six light responsive elements and six inducible responsive elements, predict that the promoter is a fully functional start the child.Second, to clone the four5’end of the deletion promoter in order, and the length of the5promoter in turn is2036bp,1440bp,996bp,652bp and344bp.5’ promoter containing the GUS reporter gene plasmid pCAMBIA1301to connect and build out of the5’end of the deletion promoter vectors.Third, all plants will be built by Agrobacterium-mediated expression vector was transformed into rice,34transgenic plants, hygromycin detection of transgenic plants, of which21tested positive. GUS staining detected positive for genetically modified organizations, according to the staining results obtained, and GUS Gene were not in the organization was not found to express.