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应用套叠PCR技术构建番木瓜eIF4E和eIFiso4E基因的hpRNA植物表达载体

Construction of hpRNA Expression Vectors of Papaya eIF4E and eIFiso4E Gene by Overlap Extension PCR
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摘要 植物真核翻译起始因子4E(eIF4E)在蛋白质合成的起始中发挥重要作用,参与植物-病毒互作,影响病毒的侵染过程.为了研究eIF4E在植物病毒侵染中的功能,建立了一种快速的套叠PCR新方法,成功构建了番木瓜eIF4E和eIFiso4E基因的hpRNA结构,并将其连接到改造的植物表达载体pBI121上,为利用RNA干扰技术研究番木瓜eIF4E和eIFiso4E基因在病毒侵染中的作用奠定了基础. Plant eukaryotic translation initiation factor 4E（eIF4E）plays an important role in the initiation of protein synthesis,which is also involved in plant-virus interactions and affects the virus infection process.To study the functions of eIF4E in plant-virus interaction,a rapid one-step and cost-efficient overlap extension PCR（OE-PCR）was developed to construct the hpRNA expression vectors of papaya eIF4E and its isomer eIFiso4E genes for RNA interference experiments.Our results suggest that this novel OE-PCR is a useful strategy for constructing hpRNA.It laid the foundation for studying the roles of papaya eIF4E and eIFiso4E genes in viral infection by RNA interference technology.
作者 吴金燕 言普 沈文涛 黎小瑛 周鹏
机构地区 中国热带农业科学院热带生物技术研究所农业部热带作物生物技术重点开放实验室 海南大学
出处 《生命科学研究》 CAS CSCD 2010年第5期393-397,共5页 Life Science Research
基金 国家自然科学基金资助项目(30760134 30960218) 海南省自然科学基金资助项目(808186) 中央级公益性科研院所基本科研业务费专项(ITTBZD07-32) 海南大学热带作物种质资源保护与开发利用教育部重点实验室开放课题
关键词 HPRNA 植物表达载体 套叠PCR eIF4E基因 eIFiso4E基因 hpRNA plant expression vector overlap extension PCR eIF4E gene eIFiso4E gene
分类号 Q782 [生物学—分子生物学]
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参考文献19

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生命科学研究
2010年 第5期

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