| 雨生红球藻八氢番茄红素合成酶基因的克隆及表征 |
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| 引用本文： | 梁成伟,赵方庆,秦 松,檀琮萍,魏 炜,孟春晓.雨生红球藻八氢番茄红素合成酶基因的克隆及表征[J].生物化学与生物物理进展,2006,33(9):854-860. |
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| 作者姓名： | 梁成伟 赵方庆 秦 松 檀琮萍 魏 炜 孟春晓 |
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| 作者单位： | 1. 中国科学院海洋研究所,青岛,266071;中国科学院研究生院,北京,100039
2. 中国科学院海洋研究所,青岛,266071 |
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| 基金项目： | 国家自然科学基金;中国科学院知识创新工程项目 |
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| 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.
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| 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 |
| 收稿时间： | 3/7/2006 10:05:28 AM |
| 修稿时间： | 5/8/2006 7:12:21 PM |
Molecular Cloning and Characterization of Phytoene Synthase Gene From a Unicellular Green Alga Haematococcus pluvialis |
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| LIANG Cheng-Wei,ZHAO Fang-Qing,QIN Song,TAN Cong-Ping,WEI Wei and MENG Chun-Xiao.Molecular Cloning and Characterization of Phytoene Synthase Gene From a Unicellular Green Alga Haematococcus pluvialis[J].Progress In Biochemistry and Biophysics,2006,33(9):854-860. |
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| Authors: | LIANG Cheng-Wei ZHAO Fang-Qing QIN Song TAN Cong-Ping WEI Wei and MENG Chun-Xiao |
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| Institution: | Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Graduate University, The Chinese Academy of Sciences, Beijing 100039, China;Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Graduate University, The Chinese Academy of Sciences, Beijing 100039, China;Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Graduate University, The Chinese Academy of Sciences, Beijing 100039, China;Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Graduate University, The Chinese Academy of Sciences, Beijing 100039, China;Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences, Qingdao 266071, China;Graduate University, The Chinese Academy of Sciences, Beijing 100039, China |
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| Abstract: | 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. pluvialis. The cDNA and genomic genes of phytoene synthase, i.e. psy from H.pluvialis were cloned and characterized.Result showed that psy had one open reading frame of 1 200 bp encoding a putative polypeptide of 400 amino acids which was interrupted by four introns. Phylogenetic analysis revealed that psy from 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 cis-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 attemptedintroducing the construct into the green alga via particle bombardment resulted in lacZ transient expression. |
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| Keywords: | Haematococcus pluvialis phytoene synthase phylogenetic analysis promoter |
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