文献类型： 外文期刊
作者： Yan, Pu 1 ; Wang, Shuchang 1 ; Shen, Wentao 1 ; Gao, Xinzheng 2 ; Wu, Jinyan 1 ; Zhou, Peng 1 ;
作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Key Lab Trop Crop Biotechnol, Minist Agr, Haikou 571101, Peoples R China
2.Hainan Med Coll, Dept Basic Med Sci, Haikou 571101, Peoples R China
关键词： virus resistance
期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）
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年卷期：
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收录情况： SCI
摘要： RNA silencing has been adopted to develop virus-resistant plants through expression of virus-derived hairpin RNAs. Due to the high sequence specificity of RNA silencing, this technology has been limited to the targeting of single viruses. Simultaneous targeting of multiple viruses or plant genes can be achieved by using a chimeric cassette. In this study, a simple method was developed to construct chimeric hairpin RNA rapidly and efficiently. This method splices two DNA fragments from viruses or plant genes to be a chimeric sequence using Overlap Extension PCR (OE-PCR); then this chimeric sequence was assembled with an intron sequence to generate an intron-containing hairpin RNA construct in one step mediated by OE-PCR. This method is neither dependent on restriction enzymes nor requires expensive consumables, so a chimeric hairpin RNA can be constructed rapidly and costlessly. Two chimeric hairpin RNA constructs were amplified successfully using this method, with the targeting sequences from both papaya ringspot virus (PRSV) and two plant genes encoding translation initiation factors elF4E and elFiso4E. This novel method is a useful strategy to construct chimeric hairpin RNA for RNA silencing in plants.
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