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Discovery of two novel miRNAs from the Ovis aries by a combinatorial approach of experiments and bioinformatics

DOI: 10.18805/ijar.B-721    | Article Id: B-721 | Page : 1155-1161
Citation :- Discovery of two novel miRNAs from the Ovis aries by a combinatorial approach of experiments and bioinformatics.Indian Journal of Animal Research.2018.(52):1155-1161
Weihua Chang, Juanhong Wang, Yong Zhang and Junyuan Wu changweihua112@126.com
Address : College of Animal Science, Tarim University, Alar City, Xinjiang 843300, China
Submitted Date : 5-03-2017
Accepted Date : 14-10-2017

Abstract

n the study, we successful constructed a library from the ovine testis using next-generation sequencing technology, and identified 219 novel miRNAs by bioinformatics. Two of the novel miRNAs (ovis_aries_testis-m0052_5p and ovis_aries_testis-m0165_5p), which were expressed in the sheep testis and ovary, and were confirmed by real-time PCR and northern blotting. Ovis_aries_testis-m0052_5p was 23 nucleotides in length, was located on chromosome 15, and had 100% similarity to mmu-miR-34c-5p, hsa-miR-34c-5p, gga-miR-34c-5p, and cfa-miR-34c. Ovis_aries_testis-m0165_5p was 21 nucleotides in length, located on chromosome 5, and had 100% similarity to mmu-miR-145a-5p, hsa-miR-145-5p, ssc-miR-145-5p, and bta-miR-145. The pre-miRNAs for Ovis_aries_testis-m0052_5p and Ovis_aries_testis-m0165_5p were 75 and 81 nucleotides in length, and both had a standard hairpin stem-loop structure. From the consistency of the sequence and structure, we speculated that ovis_aries_testis-m0052_5p had a function similar to hsa-miR-34c-5p, mmu-miR-34c-5p, and ovis_aries_testis-m0165_5p had a function similar to hsa-miR-145-5p, which were involved in the fine regulation of cell survival, spermatozoon generate, breeding activities. Therefore, we defined the identified miRNAs as oar-miR-34c-5p and oar-miR-145-5p. The results will enrich the miRNA database, and provide the basis for research into the regulatory mechanisms of miRNA in relation to breeding activities.

Keywords

MicroRNA Northern blotting Ovary Ovine Real-time PCR Testis

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