Studying For The Structure And Function Of Rice Blast Gene Pib Promoter Region By Rice Transformation

As a important component under the regulation level, promoter controls the gene expression in special organ, development phase and environmental condition. Pib gene is the first major blast resistant gene cloned from rice using map-based clone technique and belongs to NBS-LRR resistant disease gene family. Previous studies show that the expression of pib gene was induced by environment and chemistry factors. It has not yet any studies about if the expression particularity of pib gene has connected with the upstream regoins of pib structure gene or if promoter activity and the induced response components in it will affect the gene expression. So it will offer important theoretical basis for gene expression-regulation studies and resistant disease breeding that separate and identify pib gene promoter and understand its expression speciality and action mechanism. In this paper, we report initial analysis results to the promoter activity and inducement speciality of the upstream regoin of pib gene.1. Sequence analysis and restriction endonuclease digestment find that pib gene was positive-ligated into the site Not I in vector pBluescript SK(+).Pib gene includes four introns and three extrons and 3.6 kb region upstream to ATG. By analyzing pib gene promoter using PLACE we find that there are conserved components and kinds of inducement-response components upstream or downstream to ATG.2. Different regions of pib gene promoter obtained by digestion or PCR were ligated into the binary vector pCAMBIA1301 to substitute the CaMV35S promoter which is upstream to gus. The recombined plasmids:pNAR601,pNAR602,pNAR603 and pNAR604 were then constructed. All recombined binary vectors were confirmed by restricted enzyme digestion and transfered into Agrobacterium tumefaciens strain EHA101 by freeze-thaw method.