雨生红球藻八氢番茄红素合成酶基因的克隆及表征(英文)  

Molecular Cloning and Characterization of Phytoene Synthase Gene From a Unicellular Green Alga Haematococcus pluvialis

作　　者：梁成伟[1] 赵方庆[1] 秦松[1] 檀琮萍[1] 魏炜[1] 孟春晓[1] 

机构地区：[1]中国科学院海洋研究所

出　　处：《生物化学与生物物理进展》2006年第9期854-860,共7页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30470156);中国科学院创新项目(KZCX3-SW-215)~~

摘　　要：雨生红球藻是一种单细胞绿藻,在多种逆境胁迫条件下能够大量合成并迅速积累虾青素,其积累量最高可达细胞干重的4%,从而成为目前最理想的天然虾青素合成工具.八氢番茄红素合成酶(PSY)是虾青素合成途径中第一个限速酶.分离了八氢番茄红素合成酶基因(psy)的全长cDNA及基因组DNA.其全长cDNA包括1200个碱基,编码400个氨基酸,基因组DNA包括5个外显子,4个内含子.系统发育分析结果显示,绿藻的八氢番茄红素合成酶基因形成一个进化枝,它们与高等植物的psy亲缘关系比较近.通过GenomeWalking的方法,分离了psy基因约1kb的5′侧翼序列.将含有TATA-box和CAAT-box的297bp的序列与LacZ报告基因构成嵌合的表达载体,用基因枪法转化雨生红球藻.lacZ的瞬间表达检测结果表明,这段上游序列能够驱动lacZ表达,具有启动子活性.The unicellular green alga Haematococcus pluvialis accumulates a highly valuable ketocarotenoid, i.e. astaxanthin up to 4% dry weight under stress conditions. Phytoene synthase is considered to be the first rate limiting enzyme in carotenoid biosynthesis pathway in H. pluvial/s. The cDNA and genomic genes of phytoene synthase, i.e. psy fi＇om H.pluvialis were cloned and characterized. Result showed that psy had one open reading flame of 1 200 bp encoding a putative polypeptide of 400 amino acids which was interrupted by four introns. Phylogenetic analysis revealed that psy fi＇om green algae formed a monophyletic clade, and its closer relationship was higher plants. By using genomic walking approach, an approximate 1 kb 5＇ flanking region ofpsy gene was cloned and a number of putative c/s-regulatory elements were revealed. Fusing a 297 bp internal sequence （-297 to -1 bp from the translation initiation codon ofpsy） with the reporter gene, i.e. lacZ before attempted introducing the construct into the green alga via particle bombardment resulted in lacZ transient expression.

关 键 词：雨生红球藻 八氢番茄红素合成酶 5’侧翼序列 系统发育分析 启动子 

分 类 号：Q78[生物学—分子生物学]