项目名称： 版纳微型猪近交系生精障碍关键基因pSeptin12、14的功能与作用机制研究

项目编号： No.31460580

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 霍金龙

作者单位： 云南农业大学

项目金额： 50万元

中文摘要： 版纳微型猪近交系是世界上首个培育成功的大型哺乳动物近交系，培育期间出现的不育公猪致多个亚系断代，影响群体扩繁。前期我们对近交系不育和可育公猪睾丸的转录组测序及表达分析发现Septin基因家族的pSeptin12和pSeptin14在可育公猪睾丸中高表达在不育公猪中弱表达，且呈睾丸特异性和年龄依赖性，提示其可能在精子发生中发挥重要作用。本研究拟从DNA水平检测不育和可育公猪中pSeptin12、14基因的结构差异，寻找功能SNP位点并通过细胞功能实验进行验证，推测其在不育和可育公猪间差异表达的分子机制；利用免疫印迹、酵母双杂交等技术从蛋白水平研究其表达规律，鉴定与其互作的蛋白；运用RNAi技术干扰pSeptin12、14在睾丸支持细胞中的表达，然后通过RNA-seq分析差异表达基因，揭示其在精子发生中可能的信号通路及调控机制，为阐明版纳微型猪近交系生精障碍机理提供理论依据，并提出育种指导方案。

中文关键词： 版纳微型猪近交系；生精障碍；septin；功能研究；分子机制

英文摘要： Banna Mini-pig Inbred Line (BMI) is the first successfully cultivated large mammal inbred line in the world. The appearing of sterility in boar during the period of cultivation have caused several sublines to terminate and obstructed BMI population multiplication. The fact that the expression of Septin12 and Septin14 of Septin gene family member were testis-specific and age-dependent and high in the fertile boar while low in infertile boar founded by transcriptome sequencing and express analysis of infertility and fertility boar testicles suggested that Septin12 and Septin14 genes may play an important role in spermatogenesis. In this study, we intends to detect structural differences of Septin12 and 14 gene between the infertility and fertile boar on DNA level, look for the functional SNP loci and conduct functional verification via cell experiment. Further, we can speculate the molecular mechanisms for different expression between the infertility and fertile boar. We will study the expression pattern and identify interacting proteins of Septin12 and Septin 14 on protein level by western blot and yeast two-hybrid technology. We will interfer the expression of Septin12 and Septin 14 in Sertoli cells by RNAi technology and analyze the differentially expressed genes by RNA-seq in order to reveal their signaling pathways and possible regulatory mechanisms in spermatogenesis. These study will provide a theoretical basis for clarifying the spermatogenesis failure mechanism and propose a breeding guidance for BMI.

英文关键词： Banna minipig inbred line(BMI);spermatogenesis dysfunction;septin;function study;molecular mechanism