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  2018, Vol. 26 Issue (2): 234-245    DOI:
Articles and Letters Current Issue | Archive | Adv Search |
Target Regulation of the Expression of Steroidogenic Acute Regulatory Protein Gene in Sheep (Ovis aries) Ovaries by oar-mir-150
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Abstract  In order to study the regulation of Turpan black sheep (Ovis aries) in breeding and cultivate the high fecundity breeds of sheep, ovis aries microRNA-150(oar-mir-150), which was significantly differently expressed in follicular and luteal phase of Turpan black sheep, was studied as the research object in this study. The target genes of oar-mir-150 were predicted by bioinformatics, and the expression of oar-mir-150 and steroidogenic acute regulatory protein gene(STAR) in follicular and luteal phase of Turpan black sheep ovaries were detected by qRT-PCR. In addition, the base sequence of wild, mutant and deletant-type STAR gene were synthesized and ligated with the PGL3-control vector after annealing for constructing the luciferase reporter gene vector. Then, the oar-mir-150 mimic or negative control mimic with the recombinant plasmid were co-transfected into 293T cells, and the luciferase activity was detected by the double luciferase activity assays. The results indicated that oar-mir-150 had a total number of 540 target genes, and 8 target genes were differently expressed between follicular and luteal phase. STAR gene as a target gene was up-regulated when the follicular phase compare to luteal phase that had the target gene binding site (UUGGGAG) of the oar-mir-150 which located in the 3'UTR. The relative quantitative results illustrated that the relative expression of oar-mir-150 was 0.73 which is the significant decrease (P<0.05) and STAR mRNA is 1.90 which is the significantly increase(P<0.05) in the Turpan black sheep ovaries when the follicular phase compare to the luteal phase. The wild, mutant and deletant-type luciferase reporter gene vectors were all successfully constructed. The transfection ratio of transfection reagent and green fluorescent protein vector was 1∶1 which was determined as the best transfection efficiency and the transfection ratio of PGL3-control and PRL-TK was 200:1 which was determined as the optimal transfection ratio. The double luciferase activity assays demonstrated that oar-mir-150 effectively negatively regulated the expression of STAR. Studies have shown that there was a definite relationship of target regulation between oar-mir-150 and STAR, and oar-mir-150 regulated the expression of STAR gene negatively, which would provide a theoretical basis for the study of microRNA in regulating the reproduction process of Turpan black sheep at the different stages of estrus cycle.
Key wordsTurpan black sheep, Steroidogenic acute regulatory protein gene, Ovis aries microRNA -150(oar-mir-150), Estrus cycle     
Received: 05 May 2017      Published: 04 February 2018
ZTFLH:  s826  
  s813.3  
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Articles by authors
MIAO Yan-Beng
LIU Re-Nan
WEI Pan-Hui
ZHOU Rong-Yan
LI Xiang-Yun
XI Jian-Zhong
ZHANG Zhen-Hong
Cite this article:   
MIAO Yan-Beng,LIU Re-Nan,WEI Pan-Hui, et al. Target Regulation of the Expression of Steroidogenic Acute Regulatory Protein Gene in Sheep (Ovis aries) Ovaries by oar-mir-150[J]. , 2018, 26(2): 234-245.
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http://journal05.magtech.org.cn/Jwk_ny/EN/     OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2018/V26/I2/234
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