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| 早酥梨病程相关基因非表达子1基因（NPR1）克隆及原核表达 | |
| 引用本文： | 张范,张朝红,徐炎,王跃进.早酥梨病程相关基因非表达子1基因（NPR1）克隆及原核表达[J].农业生物技术学报,2010,18(1):18-23. |
| 作者姓名： | 张范 张朝红 徐炎 王跃进 |
| 作者单位： | 1. 西北农林科技大学园艺学院,杨凌,712100 2. 农业部西北园艺植物种质资源利用重点开放实验室,杨凌,712100 3. 陕西省农业分子生物学重点实验室,杨凌,712100 |
| 基金项目： | 陕西省重大科技专项，科技部863计划项N |
| 摘 要： | 本研究以成年早酥梨(Pyrus bretschneideri cv.Zaosu)叶片为材料,通过RT-PCR克隆早酥梨病程相关基因非表达子1基因(NPR1)并获得其全长序列1771 bp,开放阅读框为1761 bp,编码586个氨基酸(GenBank登录号:FJ769372).利用NCBI/Blastp和ClustalX软件进行相似性分析表明,目的基因编码蛋白质序列与日本梨(p.pyrifolia)、秋子梨(P.ussuriensis)、苹果(Malus xdomestica)、烟草(Nicotiana tabacum)和拟南芥(Arabidopsis thaliana)的NPR1蛋白相似性分别为99%、98%、98%、67%和59%.将其连接pGEX-4T-1原核表达载体并转化大肠杆菌(EScherichia coli)BL21,经IPTG诱导表达获得大小约为91 kD的目的融合蛋白,融合蛋白主要以包涵体形式表达,表达量约占总蛋白17%. |
| 关 键 词： | 早酥梨 病程相关基因非表达子1基因(NPRl) 克隆 原核表达 |
Cloning and Prokaryotic Expression of Zaosu Pear Nonexpressor of Pathogenesis-related Genes 1 Gene(NPR1) in Escherichia coli |
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| Zhang Fan Zhang Chaohong Xu Yan Wang Yuejin Northwest Agriculture , Forestry University,Yangling, Key Laboratory of Northwest Horticulture Plant Germplasm Utilization,Ministry of Agriculture, Key Laboratory of Agricultural Molecular Biology of Shaanxi Province,College of Horticulture.Cloning and Prokaryotic Expression of Zaosu Pear Nonexpressor of Pathogenesis-related Genes 1 Gene(NPR1) in Escherichia coli[J].Journal of Agricultural Biotechnology,2010,18(1):18-23. | |
| Authors: | Zhang Fan Zhang Chaohong Xu Yan Wang Yuejin Northwest Agriculture Forestry University Yangling Key Laboratory of Northwest Horticulture Plant Germplasm Utilization Ministry of Agriculture Key Laboratory of Agricultural Molecular Biology of Shaanxi Province College of Horticulture |
| Institution: | Zhang Fan Zhang Chaohong Xu Yan Wang Yuejin * 1 Northwest Agriculture , Forestry University,Yangling,712100*,2 Key Laboratory of Northwest Horticulture Plant Germplasm Utilization,Ministry of Agriculture,712100,3 Key Laboratory of Agricultural Molecular Biology of Shaanxi Province,College of Horticulture |
| Abstract: | The total RNA was extracted from leaves of Zaosu pear(Pyrus bretschneideri cv.Zaosu).Using RT-PCR method,the full-length cDNA of nonexpressor of pathogenesis-related genes 1 gene(NPR1) was cloned from the Zaosu pear leaves.Bioinformatics analysis showed that the obtained 1 771 bp fragment contained a 1 761 bp open reading frame encoding 586 amino acid residues(GenBank accession No.FJ769372).The homology rates of amino acid sequences of NPR1 gene between Zaosu pear and P.pyrifolia(ABA62792),P.ussuriensis,Mal... |
| Keywords: | Zaosu pear Nonexpressor of pathogenesis-related genes 1 gene(NPR1) Cloning Prokaryotic expres sion |
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