Rapid one-step construction of hairpin RNA

文献类型： 外文期刊

第一作者： Yan, Pu

作者： Yan, Pu;Shen, Wentao;Gao, Xinzheng;Zhou, Peng;Duan, Jun

作者机构：

关键词： RNAi;hpRNA;OE-PCR;Asymmetric PCR;Plant;PRSV;Antivirus

期刊名称：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS （ 影响因子：3.575； 五年影响因子：3.381 ）

ISSN： 0006-291X

年卷期： 2009 年 383 卷 4 期

页码：

收录情况： SCI

摘要： Hairpin RNA (hpRNA) is commonly used for gene-function exploration and gene engineering. In this study, a novel method was developed to construct intron-containing hairpin RNA (ihpRNA) rapidly and efficiently based on Overlap Extension PCR (OE-PCR). This method, Mixed One-step OE-PCR (MOOE-PCR), can amplify two inverted repeats of DNA fragments and a spliceable intron in parallel, and then assemble them to generate ihpRNA constructs in the same tube without the purification of intermediate products. This method required a PCR process of 38-40 cycles and ordinary PCR reagents. A total of 10 ihpRNA constructs were amplified successfully using this method, with the stems ranging from 50 bp to 484 bp in length, Our results suggest that this novel method is a useful strategy for constructing ihpRNA. (C) 2009 Elsevier Inc. All rights reserved.

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