Screening And Analysis For Differential Expressed Proteins In Ewes Ovary Tissue Of Different Lambing Performance

Lambing trait is one of the most important economic traits in sheep,There are three lambing Performances: singles births, twins births and multiple births, Lambing rate in different breeds have quite differences. The most effective way of breeding sheep lambing traits of the ideal type is finding lambing traits of major genes and molecular markers, and developing molecular breeding from sheep genetic substance DNA molecules. Sheep lambing number is beyond the control of ovulation, during the process of follicular development and ovulation, there are a series of cellular regulatory mechanisms, involving various types of protein-protein interactions Therefore, if we need to have a comprehensive and in-depth investigating for complex regulation of metabolic pathways of sheep lambing performance, we need study in integrated, dynamic and network level to study on differential proteins in sheep lambing performance, identify major genes and transmission pathways, reveal molecular genetic mechanisms and regulation mechanism to provide the theory for molecular markers and molecular breeding of sheep lambing performance.In this study, sheep ovarian protein optimum protein extract method was first selected and optimized, and then composition and content of sheep ovarian protein for Mongolian ewes with singles, Mongolia ewes with twins, Dorset ewes with singles,Dorset ewes with twins and Small Tail Han ewes with multiple births were isolated and identified by 2-DE and MS, and acquired the differential proteins. Last, the bioinformatics was used to analyze for differential proteins in Cellular Component,molecular function, biological process, KEGG Pathways and PPI to explore the molecular mechanism and regulation mechanism of sheep lambing performance. The main results were as follows:⑴ In the study, TCA/acetone precipitation and direct lysis methods were used to extract the sheep ovarian protein, and then to optimize the direct lysis methods, it could get more spots and veracity while extracting more small molecule-protein with direct lysis methods by 2-DE for different extraction methods of sheep ovarian protein,and the optimal prescription of direct lysis methods was 7M urea, 2M thiourea, 4%CHAPS, 0.5%-amphoteric electrolyte p H 3-10, 100 mmol/l DTT, 60 mmol/L Tris,20μl protease inhibitor cocktail.⑵ The Target P software was used for subcellular localization of different proteins. 2 proteins were present in the secretory pathway, 3 proteins were present in Mitochondria, 14 proteins were present in other pathway.The BLAST2 GO software was used for Gene Ontology analysis, there were 21 commentary of 19 proteins in Cellular Component, and all commentary of enrichment were cytoplasm, cytosol, nucleus and calcium complex, there were 31 commentary of18 proteins(except for nitric oxide-inducible gene protein) in molecular function, and all commentary of enrichment were ATP binding, signal transducer activity, calcium ion binding, oxygen transporter activity, growth factor activity, DNA binding and carbohydrate binding, there were 35 commentary of 18 proteins(except for phosphatidylethanolamine-binding protein 1-like) in biological process, and all commentary of enrichment were oxidation-reduction process, ovarian follicle development, signal transduction, BMP signaling pathway, growth factor receptor signaling pathway, C21-steroid hormone metabolic process and metabolic process.The KAAS database was used for KEGG analysis of different proteins, there were 23 related Pathways of 13 proteins, and all Pathways of enrichment were Oxidative phosphorylation pathway, Oocyte meiosis, TGF-beta signaling pathway and Gn RH signaling pathway.⑶ Calmodulin, GDF9, BMPR-1B, MTHFR and SOD-[Cu-Zn]-like were elected to the candidate proteins of sheep lambing performance.