Identification of a novel miRNA from the ovine ovary by a combinatorial
approach of bioinformatics and experiments |
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Authors: | Weihua CHANG Juanhong WANG Dayong TAO Yong ZHANG Jianzhong HE Changqing SHI |
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Institution: | 1)College of Animal Science, Tarim University, Alar City, Xinjiang 843300, PR China;2)College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, Gansu 730070, PR China |
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Abstract: | MicroRNAs (miRNAs) are a class of short endogenous, single-stranded, non-coding small RNA
molecules, about 19–25 nucleotides in length that regulate gene expression at the
translation level and influence many physiological process, such apoptosis, metabolism,
signal transduction, and occurrence and development of diseases. In this study, we
constructed a library from the ovine luteal phase ovary by using next-generation
sequencing technology (Solexa high-throughput sequencing technique) and identified 267
novel miRNAs by bioinformatics. One of the novel miRNAs (ovis_aries_ovary-m0033_3p), which
expressed in the sheep ovary and testis, was confirmed by real time PCR and northern blot.
Ovis_aries_ovary-m0033_3p was 21 nucleotides in length and located on chromosome 12, and
it had 100% similarity to hsa-miR-214-3p, mmu-miR-214-3p, dre-miR-214and ssc-miR-214.
Meanwhile, the pre-miRNA was 82 nucleotides in length and had a standard hairpin stem-loop
structure. From the consistency of the sequence and structure, we speculated that
ovis_aries_ovary-m0033_3p had a function similar to hsa-miR-214-3p, which is involved in
the fine regulation of cell survival, embryonic development, breeding activities and
resistance to ovarian cancer, so we defined it as oar-miR-214-3p. These experimental
results will enrich the miRNA database for ovis aries and provide the
basis for researching the regulation mechanism of miRNA in relation to breeding activities
of seasonal breeding animals. |
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Keywords: | microRNA next-generation sequencing northern blot ovine real-time PCR |
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