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欧李PSY基因cDNA克隆与表达分析

发表时间:2013-10-15  浏览量:1295  下载量:524
全部作者: 张建成,王鹏飞,刘和,薛晓芳,穆晓鹏,杜俊杰
作者单位: 山西农业大学园艺学院
摘 要: 利用GenBank登录的植物八氢番茄红素合成酶(phytoene synthase,PSY)氨基酸保守区序列设计简并引物,通过反转录PCR(reverse transcribed-PCR,RT-PCR)和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)从欧李[Cerasus humilis(Bge)Sok]果实中获得了一个PSY cDNA全长序列,命名为ChPSY. 序列分析表明:该基因cDNA全长1 559 bp,最大开放读码框为1 194 bp,编码397个氨基酸,与草莓PSY氨基酸序列的同源性为90.4%,与辣椒、拟南芥、柑橘、番木瓜、玉米和水稻等植物的同源性为65.7%~72.5%. 半定量RT-PCR分析表明:ChPSY在欧李的不同组织中均有表达,表达量依次为花>深红果>橙红果>泛白果>青绿果>老叶>幼叶。工程大肠杆菌异源表达体系证实ChPSY可编码一个功能蛋白,催化工程菌株中2分子r{牛儿基r{牛儿基焦磷酸(geranylgeranylpyrophosphate,GGPP)缩合生成八氢番茄红素。
关 键 词: 果树学;欧李;八氢番茄红素合成酶;cDNA克隆;表达分析
Title: Cloning and expression analysis of PSY gene from Cerasus humilis (Bge) Sok
Author: ZHANG Jiancheng, WANG Pengfei, LIU He, XUE Xiaofang, MU Xiaopeng, DU Junjie
Organization: College of Horticulture, Shanxi Agricultural University
Abstract: Based on the conserved amino acid sequences of phytoene synthase (PSY) from known plant in GenBank, the PCR primers were designed to amplify cDNA fragments from the fruit of Cerasus humilis (Bge) Sok by reverse transcribed-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE), a PSY gene named ChPSY was cloned. Sequence analysis indicated that the cDNA full-length was 1 559 bp, which contained an open reading frame of 1 194 bp and encoded a protein of 397 amino acid residues. The ChPSY protein showed 90.4% homology to the PSY from strawberry, and also 65.7%-72.5% homology to the PSYs from other plants, such as pepper, Arabidopsis thaliana, melo, papaya, maize and rice. The semi-quantitative RT-PCR analysis revealed that expression of ChPSY could be detected in different tissues of C. humilis. The expression level from high to low in the following order: flower> deep-red fruits> reddish-orange fruits>white fruis>green fruits>old leaf>new leaves. The heterogenous expression in Escherichia coli system confirmed that ChPSY could encode a functional phytoene synthase which could catalyze a condensation of two molecular of geranylgeranylpyrophosphate (GGPP) to form phytoene in an engineering E. coli.
Key words: pomology; Cerasus humilis(Bge)Sok; phytoene synthase; cDNA cloning; expression analysis
发表期数: 2013年10月第19期
引用格式: 张建成,王鹏飞,刘和,等. 欧李PSY基因cDNA克隆与表达分析[J]. 中国科技论文在线精品论文,2013,6(19):1830-1839.
 
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