title: Computational identification of microRNAs in peach expressed sequence tags and validation of their precise sequences by miR-RACE.
abstract: 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 5' and 3' 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.
authors.affiliation: College of Horticulture, Nanjing Agricultural University, Nanjing 210095, People's Republic of China. 2009104022@njau.edu.cn
authors.forename: Yanping
authors.initials: Y
authors.lastname: Zhang
authors.forename: Mingliang
authors.initials: M
authors.lastname: Yu
authors.forename: Huaping
authors.initials: H
authors.lastname: Yu
authors.forename: Jian
authors.initials: J
authors.lastname: Han
authors.forename: Changnian
authors.initials: C
authors.lastname: Song
authors.forename: Ruijuan
authors.initials: R
authors.lastname: Ma
authors.forename: Jinggui
authors.initials: J
authors.lastname: Fang
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D020224:Expressed Sequence Tags; D035264:Flowers / Q000378:metabolism; D005638:Fruit / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D018515:Plant Leaves / Q000378:metabolism; D027861:Prunus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D035683:MicroRNAs