Cloning, Characterization And Expression Identification Of Goat Ovarian MiRNAs

There are some problems such as lack of high fecundity breed and lower lambing rate in our country sheep industry. The key to solve these problems need to explicate the mechanism of goat ovary, raise lambing rate, foster new breed. Although researchers had got many breakthrough on reproductive performance of goat in past40years, there are lots of problems need to be researched in-depth. The mechanism of follicle development, select, ovulate of goat is not clear, the factor of majority ovulate is also unclear. Ovary is a cyclical upgrowth tissue^follicle generate、mature、hormone synthesize and ovulate all be controlled by many factors. microRNAs (miRNAs) is a small endogenous non-coding RNA molecules, contain21-25nucleotide, acting on a or several target mRNAs, and then to exert post-transcriptional control by target mRNA scission or suppression. miRNAs which expressed in ovary cell play a key role in the regulation of genes expression during follicles growth. But these studies always centralize on rat or mouse, goat follicular development and ovulation mechanism are different with mice. It’s necessary to ascertain goat ovary miRNA herd and identify expression of tissue cell against these question, which can replenish and consummate the mechanism of goat reproduction performance. Meantime it can also provide important reference value of treatment of ovarian diseases.Consequently, our study use Dazu black goat ovary and Inner Mongolia cashmere goat ovary as experiment materies. In the first place,adopting High-throughput sequencing to determine miRNAs in goat ovary cell; and then using qPCR to detect the expression of21miRNAs which have higher read count than the rest, finally,taking advantage of bioinformatics technology to predict the target genes of miRNAs. The main resμLts are as following:1. We got80known miRNAs and108new miRNAs by constructing Dazu black goat ovaries and Inner Mongolia cashmere goat ovarian mixed library. There are155miRNAs belonging to89miRNA families,33miRNAs do not belong to any family.2. We obtained14important target genes with fecundity by software of PITA which specially predict target genes of miRNAs.3. Goat ovaries and other organizations expression profiling were determined by Fluorescence quantitative qPCR technology. There are10higher relative expression of miRNAs (chi-miR-2209d-3p、chi-miR-2567d-5p、chi-miR-4465a-5p、 chi-miR533g-3p、chi-miR-100f-5p、chi-miR-M1-16-5p、chi-miR-1491a-5p、 chi-miR-152c-3p、miR-191-5p and miR-241a-5p) in two goat ovaries. chi-miR-2209d-3p denote significant differences in the expression profiles of both ovarian was calculated.