Kas基因启动子区的克隆及功能初步分析

Clone and Primary Functional Analysis of Promoter Region of 3-oxoacyl-[acyl-carrier-protein] Synthase(Kas) Gene

利用TAIL-PCR技术,克隆到了与辣椒素合成有关的胎座特异表达基因———3-酮酯酰-ACP合成酶基因(Kas)上游400 bp的调控区域.将其全长片段与GUS基因连接构建植物表达载体并转化烟草.GUS组织化学染色表明,克隆到的440 bp片段具有启动子活性.对该片段进行序列分析发现,在起始密码子ATG上游存在2个TATA-box,分别为-316^-311位的TATAAA和-224^-219位的TATAAA;在TATA-box上游还存在1个位于-378^-374处的CAAT-box,序列为CCAAT.该研究旨在为利用基因调控辣椒素的生物合成,提高辣椒果实中的辣椒素含量奠定基础.

A 440 bp length of DNA fragment was cloned from upstream region of 3-oxoacyl-[ acyl-carrier- protein] synthase （Kas） gene, a placental-specific gene encoding capsaicinoids biosynthetic activities using TAIL-PCR strategy. A plant expression vector was then constructed by ligating the cloned fragment to GUS gene, used for transformation of tobacco. GUS staining of transgenic tobacco leaves proved that the cloned fragment could promote expression of GUS gene. Two TATA-boxes and a CAAT-box located in the upstream region of the TATA-box were further found in the DNA sequence of the cloned DNA fragment. The results from both tobacco transformation and DNA sequence analysis suggest that the cloned 440 bp DNA fragment has a promoter activity. This research is an initial try toward clone of specific promoter and gene regulation of the capsaicinoids biosynthesis in pepper fruit.