高效发掘和鉴定版纳微型猪近交系雄性不育关键基因

版纳微型猪近交系是生物医学的理想实验动物，也是异种器官移植的良好供体，但在35年的繁育过程中有多个亚系断代，前期研究发现主要是由于一些公猪不育导致。针对这一问题，本研究利用高通量技术对相同环境条件下同月龄的近交系不育公猪和正常公猪生殖器官——睾丸及附睾开展转录组和全基因组甲基化高通量测序，结合前期近交系全基因组重测序结果和NCBI上家猪基因组数据，比较两者间的差异表达基因及差异甲基化区域，综合分析鉴定近交系雄性不育关键基因、构建生精障碍功能基因网络。进一步，采用基因克隆、qRT-PCR、RNAi、过表达、免疫荧光和Western blot等技术对关键基因进行基本特性分析和转录调控研究，阐明近交系雄性不育关键基因的表达调控机理及互作网络关系，以期为版纳微型猪近交系扩繁提供分子水平的育种指导，为男性不育的临床诊治提供参考，本研究具有重要的理论意义和应用前景。

Banna mini-pig inbred line (BMI) is generally considered to be the ideal biomedical laboratory animals and excellent donor animals in pig-to-human xenotransplantation. However, preliminary study found that there are multiple substrains dying out because of some male individuals being infertile in nearly 35 years of continuous highly inbreding. To solve this problem, this study is to compare the differentially expressed genes and differentially methylated regions between infertile and normal BMI boars from the same environment and age by analyzing their reproductive organs---testes and epididymis using high-throughput transcriptome and genome-wide DNA methylation sequencing, combined with the existing BMI genome-wide re-sequencing results and pig genome database in NCBI. Afterwards, we will comprehensively analyze to identify the key inbred male sterile genes and build network of the functional genes affecting the spermatogenic disorder. Finally, in order to clarify the expression regulation mechanism and interaction network relationship of the key inbred male sterile genes, we plan to study the basic characteristics and transcriptional regulation of key genes by gene cloning, qRT-PCR, immunohistochemistry, RNAi, overexpression, immunofluorescence and Western blot technologies. These studies will provide a breeding guide for BMI reproduction on molecular level and reference on clinical diagnosis and treatment of human male infertility, which has important theoretical significance and application prospects.