The expression of oar-miR-143 in different developmental stages of sheep ovaries and the verification of target gene CDK2
In the early stage of this study,high-throughput sequencing was used to screen out the differential expression of oar-miR-143 in different stages of ovarian development of small-tailed Han sheep,and its target genes were predicted.GO and KEGG analysis suggested that oar-miR-143 might play an important role in ovarian development.The purpose of this study was to verify the target genes pre-dicted for oar-miR-143 and explore the possible molecular mechanisms of its influence and regulation on ovarian development.The relationship between the screened oar-miR-143 and its candidate target gene CDK2 was verified by dual luciferase detection system.First,the binding sites of miRNA and its candidate target genes were predicted,and then wild-type and mutant vectors of candidate target gene CDK2 containing target sites were constructed.After co-transfection with oar-miR-143 mimics into 293T cells,dual luciferase activity was detected.The results showed that oar-miR-143 could inhibit the relative luciferase activity of wild-type CDK2 3'UTR vector,but had no effect on the relative lu-ciferase activity of mutant CDK2 3'UTR vector,indicating that oar-miR-143 could bind to the CDK2 gene seed sequence to inhibit the expression of CDK2.CDK2 was confirmed as the target genes of oar-miR-143.These data will provide an important theoretical basis for elucidating the molecular regulatory mecha-nism of ovarian development in sheep.It can provide an important theoretical basis for determining the biomarkers affecting the reproductive performance of small-tailed Han sheep.
small-tailed Han sheepovarian developmentoar-miR-143target genedual luciferase reporting system
国家科技期刊平台
NSTL
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