雨生红球藻八氢番茄红素脱氢酶pds基因上游序列的分离和分析

Cloning and functional characterization of phytoene desaturase gene (pds) promoter from Haematococcus pluvlialis

在某些蓝藻、绿藻和高等植物中,八氢番茄红素脱氢酶是β-胡萝卜素合成过程中的一个关键酶之一,应用巢式PCR方法从雨生红球藻中克隆了八氢番茄红素脱氢酶基因约1kb的5’上游序列。通过生物信息学方法进行序列分析,发现pds基因上游序列中包含了一些可能的顺式元件,如ABRE调控元件、C-repeat/DRE调控元件等。同时,构建了由pds-启动子控制报告基因的重组载体,并转化到雨生红球藻细胞中,进行了报告基因瞬间表达的检测。结果表明,克隆的pds启动子区域具有启动子活性,可以驱使报告基因进行瞬间表达。

Phytoene desaturase is one of the most important enzymes necessary for the biosynthesis of carotenoids in some cyanobacteria, green algae and plants. To locate transcriptional regulation elements, we isolated an approximate 1 kb of 5＇flanking region by genome-walking method. Results of bioinformatic analysis showed that several putative cis-elements like the ABRE motif （abscisic acid responsive element）, the C-repeat/DRE （dehydration responsive element） motif and the GCN4 motif were located in 5＇flanking region ofpds. Then, we constructed the recombinant plasmid containing the reporter gene lacZ driven by pds promoter and transferred the plasmid into the cells. The transient expression results showed thatpds promoter activity can be detected.