Study Of MicroRNA In Ovis Aries Ovary Tissue

MiRNA(microRNA) is noncoding, approximately18~25bp nucleic acids widelyspreaded in eukaryote genome, regulating gene expression in translation level. Wordhad been done in identifying many miRNA, analysis of their function and the way ofdoing it has been highlighted in mulecular biology research. There is limitedinformation about Ovis aries miRNAs, and research on the miRNA associated withreproductive traits is much more scarce.In this study, Ovis aries ovary miRNA cDNA library was constructed andanalysed by direct cloning method. Real Time PCR was adopted to detect theirexpression, functions and machnisms in Ovis aries ovary. Following results areconcluded.1. Ovis aries ovary small RNA cDNA library was successfully constructed bydirect cloning methods, in which,30Ovis aries miRNA were recovered.2. Through blast the sequencein miRBase database,20miRNA sequences werehomologous to conserved miRNA,defined as Ovis aries known miRNA. In addition,oar-miR-3955-5p had not miRBase database notesin other species, possiblly specialfor Ovis aries.3. The remaining10sequences defined as Ovis aries new miRNA by secondarystructureanalysis. Although these sequences was first described in Ovis aries, andmostare highly homologous with other species miRNA. Interestingly,ovis-aries-ovary-m0001-5p only had some homology with bovine miRNAbta-miR-2285.4.10miRNAs (five known miRNA and five new miRNA) identified bycomputational method for expression analysis by Real-time PCR.All of them could bedetected inOvis aries ovary, and ovis-aries-ovary-m0003-3p had a significant higherexpression level inOvis aries ovary than the other miRNA.