Functional Analisis Of Rice Pi-hit-1 Gene Promoter

pi-hit-1 gene was a novel gene identified by using differential display PCR (DD-PCR) technique, and was predicted as a blast related gene. pi-hit-1 gene expressed higher in space flight induced blast sensitive mutant 972-4 then on-ground control 972ck. Sequence analysis revealed that pi-hit-1 gene encodes a member of ATP-dependent Clp protease family. Expression analysis revealed that of pi-hit-1 gene was induced by blast isolate, CH45. There is no different in gene structure both in 972-4 and 972ck.So it is necessary to study the transcription regularlition of pi-hit-1 gene. To further analyze the pi-hit-1 promoter structure and function, we used three database of plant gene promoter analyze software((PlantProm DB-TSSP,TFSEARCH&PLACE)) to predict the promoter of pi-hit-1 gene, we found there did have some cis-elements in about 500bp upstream of pi-hit-1 gene, also have a TATA box in 200bp upstream, then we used EMSA to analyze the promoter, the result is the same to the predict about the TATA box. To study the function of the promoter, the promoter region of pi-hit-1 from 972-4 ( 1 kb/0.5kb) was fused with the GUS gene and cloned into pCAMBIA1381Z vector. The vector was introduced to rice line TP309 with Agrobacterium-mediated transformation. The transformed callus showed no or low expression patterns.