【摘要】 为探讨HSP27基因在藏羊卵巢卵母细胞成熟及排卵过程中的作用通路，本研究构建了HSP27基因真核表达体系，利用免疫共沉淀、双向电泳及生物质谱技术鉴定藏羊卵巢全蛋白提取物中与HSP27蛋白特异性相互作用的蛋白，构建互作网络，通过生物信息学分析并预测HSP27基因在藏羊卵巢卵泡发育过程的作用通路。结果表明:藏羊卵巢全蛋白提取物中的HSP10、ERK1、Bcl-2、BMPR-1B、MTHFR、RhoD、ANXA2、Calmodulin、Transgelin、GDF5、Galectin-1、PEBP、LDH-1和SOD[Cu-Zn]等14种已识别候选蛋白与HSP27蛋白具有特异性相互作用;通过生物信息学分析表明目的蛋白在TGF-β、MAPK和m TOR信号通路中构成了复杂且紧密的交互网络，同时基于互作结果设计HSP27蛋白调节藏羊卵母细胞成熟和排卵的代谢途径，发现HSP27蛋白与BMPR-1B、GDF5、ERK1、Calmodulin、Bcl-2和SOD[Cu-Zn]具有强烈的交互作用。本研究结果为进一步探讨藏羊卵泡发育、成熟和排卵机理，以及HSP27基因作为藏羊繁殖性状目的基因用以提高藏羊产羔率提供了基础资料和理论依据。更多还原

【Abstract】 An eukaryotic expression system of HSP27 gene is constructed in order to explore the role of HSP27 gene in the maturation and ovulation process of Tibetan sheep ovary oocytes. The proteins specifically interacting with HSP27 protein in the whole protein extract of Tibetan sheep ovary are identified by immunoprecipitation,two-dimensional electrophoresis and biological mass spectrometry.The interaction network is constructed as well. The pathway of HSP27 gene in the development of ovarian follicles in Tibetan sheep is analyzed and predicted by bioinformatics. The results show that there are 14 identified candidate proteins in the whole protein extract of Tibetan sheep ovary specifically interacted with HSP27,including HSP10,ERK1,Bcl-2,BMPR-1B,MTHFR,RhoD,ANXA2,Calmodulin,Transgelin,GDF5,Galectin-1,PEBP,LDH-1 and SOD [Cu-Zn ];bioinformatics analysis shows that a complex and tight interaction network is formed by the target protein in the signaling pathways including TGF-β,MAPK and m TOR. HSP27 protein is designed to regulate the metabolic pathway of Tibetan sheep oocyte maturation and ovulation based on the interaction results. It is found that HSP27 protein shows strong interaction with BMPR-1B,GDF5,ERK1,Calmodulin,Bcl-2 and SOD[Cu-Zn]. This study provides a theoretical basis for further exploring the mechanism of follicle development,maturation and ovulation of Tibetan sheep,and HSP27 gene as the target gene of Tibetan sheep reproductive traits to improve its lambing rate.更多还原