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| 反向PCR法扩增花粉特异表达基因PSG076启动子 | |
| 引用本文： | 陈泠,苏佩佩,佟汉文,刘易科,朱展望,杨广笑,高春保,何光源.反向PCR法扩增花粉特异表达基因PSG076启动子[J].武汉植物学研究,2012,30(3):309-314. |
| 作者姓名： | 陈泠 苏佩佩 佟汉文 刘易科 朱展望 杨广笑 高春保 何光源 |
| 作者单位： | 1. 华中科技大学生命科学与技术学院中英HUST-RRes基因T程和基因组学联合实验室,科技部国际科技合作基地(基因工程),教育部分子生物物理重点实验室,武汉430074;湖北省农业科学院粮食作物研究所,粮食作物种质创新与遗传改良湖北省重点实验室,武汉430064 2. 华中科技大学生命科学与技术学院中英HUST-RRes基因T程和基因组学联合实验室,科技部国际科技合作基地(基因工程),教育部分子生物物理重点实验室,武汉430074 3. 湖北省农业科学院粮食作物研究所,粮食作物种质创新与遗传改良湖北省重点实验室,武汉430064 |
| 基金项目： | 科技部国际合作重点项目，博士后科学基金 |
| 摘 要： | PSG076基因是从奥地利小麦品种‘Ferdinand’中分离的花粉特异性表达基因,功能未知.为获得可用于小麦基因工程的花粉特异性启动子,采用优化的反向PCR法分离PSG076启动子,获得了起始密码子上游约1.4 kb的启动子序列.生物信息学分析显示,该启动子除含有与花粉特异性表达相关的调控元件AGAAA和GTGA外,还含有花粉特异性表达相关的数量元件AGGTCA和AAATGA,推测其为活性较强的花粉特异性启动子.实验中对反向PCR方法的优化可提高扩增侧翼未知序列的效率,特别适用于启动子序列的扩增. |
| 关 键 词： | 小麦 花粉特异性启动子 反向PCR 序列分析 |
Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR |
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| CHEN Ling , SU Pei-Pei , TONG Han-Wen , LIU Yi-Ke , ZHU Zhan-Wang , YANG Guang-Xiao , GAO Chun-Bao , HE Guang-Yuan.Isolation of the Promoter Region of a Pollen-specific Gene PSG076 by Inverse-PCR[J].Journal of Wuhan Botanical Research,2012,30(3):309-314. | |
| Authors: | CHEN Ling SU Pei-Pei TONG Han-Wen LIU Yi-Ke ZHU Zhan-Wang YANG Guang-Xiao GAO Chun-Bao HE Guang-Yuan |
| Institution: | 1＊(1.China-UK HUST-RRes Genetic Engineering and Genomics Joint Laboratory,The Genetic Engineering International Cooperation Base of Chinese Ministry of Science and Technology,The Key Laboratory of Molecular Biophysics of Ministry of Education,College of Life Science and Technology,Huazhong University of Science ＆ Technology(HUST),Wuhan 430074,China;2.Institute of Food Crops,Hubei Academy of Agricultural Sciences,Hubei Key Laboratory of Food Crop Germplasm and Genetic Improvement,Wuhan 430064,China) |
| Abstract: | A pollen-specific gene,PSG076,with unknown function,was isolated from Austrian wheat ’Ferdinand’.To obtain pollen-specific promoter in wheat genetic engineering,the promoter region of PSG076 gene was isolated by an improved inverse-PCR（IPCR） method.A 1.4 kb promoter sequence was cloned and analyzed by Bioinformatics tools.Some cis-acting elements or enhancers conferred to pollen-specific expression were found,such as AGAAA,GTGA,AGGTCA and AAATGA.This suggested that PSG076 promoter might be a pollen-specific promoter with strong activity.The results also showed that the improved IPCR method was efficient to isolate the flanking unknown sequence,especially in promoter isolation. |
| Keywords: | Wheat Pollen-specific promoter Inverse-PCR Sequence analysis |
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