Biotechnology Bulletin ›› 2019, Vol. 35 ›› Issue (1): 58-66.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0671

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Precursor Cloning and Tissue Expression Analysis of Yak miR-378

JI Hui1, WANG Hui1, CHAI Zhi-xin1, WANG Ji-kun1, LUO Xiao-lin2, JI Qiu-mei3, XIN Jin-wei3, ZHONG Jin-cheng1   

  1. 1. Key Laboratory of Animal Genetics and Breeding of the Ministry of Education,Southwest Minzu University,,Chengdu 610041;
    2. Sichuan Grassland Science Academy,Chengdu 611731;
    3. State Key Laboratory of Barley and Yak Germplasm Resources and Genetic Improvement,Tibet Academy of Agricultural and Animal Husbandry Sciences,Lhasa 850009
  • Received:2018-07-23 Online:2019-01-26 Published:2019-01-23

Abstract: This work aims to clone the precursor sequence of yak miR-378,to elucidate its tissue expression pattern,and to explore the regulatory function of miR-378 in the growth and development of yak combined with bioinformatics prediction and analysis of bta-miR-378 target gene. PCR was employed to successfully clone the miR-378 precursor sequence of leiwuqi yak,and real-time quantitative PCR(RT-qPCR)was used to detect the expression patterns of miR-378-3p in various tissues. Further,bioinformatics software and databases such as TargetScan,DAVID,NCBI,miRbase,etc were together applied to perform conservative analysis,target gene prediction and biological function analysis of miR-378. The results showed that miR-378 was highly conserved among different species and miR-378-3p was widely expressed in all tissues,the expression level was the highest in gluteus maximus,which was significantly higher than that in other tissues(P < 0.01). Expression in hip fat was higher than in ovary,brain,breast and liver. The obtained 272 target genes were mainly involved in many biological processes such as cell differentiation,cell development,and macromolecular metabolism,involved in progesterone-mediated oocyte maturation and gonadotropin releasing hormone(GnRH)signaling pathways. It is speculated that miR-378 may play a key role in follicular development and oocyte maturation,which may affect the reproductive performance of female calves.

Key words: yak, miR-378, cloning, tissue expression, biological information analysis