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Computational identification of microRNAs in peach expressed sequence tags and validation of their precise sequences by miR-RACE

文献类型： 外文期刊

作者： Zhang, Yanping 1 ; Yu, Mingliang 2 ; Yu, Huaping 1 ; Han, Jian 1 ; Song, Changnian 1 ; Ma, Ruijuan 2 ; Fang, Jinggui 1 ;

作者机构： 1.Nanjing Agr Univ, Coll Hort, Nanjing 210095, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Hort, Nanjing 210014, Peoples R China

关键词： Prunus persica;microRNAs;Computational prediction;qRT-PCR;miR-RACE

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

ISSN：

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收录情况： SCI

摘要： Twenty-two potential miRNAs from seven miRNA families were first predicted from more than 80,857 EST sequences of peach (Prunus persica). Using two specific 50 and 30 miRNA RACE (miR-RACE) PCR reactions and sequence-directed cloning, we accurately determined the precise sequences, especially both ends, of eight candidate miRNAs. The sequencing results demonstrated that the ppe-miRNAs were conserved to those that were predicted computationally except ppe-miR171b. We validated the existence of two members (ppe-miR171a and miR171b) of the miR171 family in peach that belonged to different precursors. qRT-PCR was further employed in analyzing expression of the eight miRNAs in peach leaves, flowers, and fruits at different developing stages, where some of the miRNAs showed tissue-specific expression.

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