无精子症DAZ基因家族在版纳微型猪近交系中的功能与作用机制研究

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，但公猪的精子质量影响了群体的扩繁，限制了其发展和应用。前期我们对无精子症DAZ家族BOLL和DAZL基因的研究发现其mRNA在睾丸中特异高表达；另外，发现BOLL起始密码子由ATG突变为了ACG，提示该基因家族在近交系精子发生过程中的作用模式可能与在其他物种中不同。针对这一发现，我们提出了三种假设，并设计实验方案，通过分析基因组成、结构及功能元件，确定BOLL基因起始密码子缺失在不同品种猪中是否具有普遍性，并揭示基因的基本特性；采用BSP-PCR、qPCR、WB、IHC技术从启动子甲基化、RNA转录以及蛋白翻译角度研究基因表达；进一步采用细胞转录组测序联合ChIP、RIP、Co-IP和RNA Pull Down进行综合分析，获悉其互作调控网络。研究结果可为阐明DAZ基因家族在版纳微型猪近交系精子发生中的功能及作用机制提供理论依据。

The poor sperm quality of the boar has limited the development and applications of Banna mini-pig inbred line (BMI), which is the first successfully cultivated large mammal inbred line in the world. Our previous study about DAZ gene family, known as deleted in azoospermia, showed that the mRNA of BOLL and DAZL has been overexpressed specifically in the testis. Besides, the initiation codon of Boll gene mutated from ATG to ACG, which indicated that the action mode of this gene family may not be exactly the same as that of other species. Hence, we proposed three different hypotheses and designed appropriate schemes to confirm the universality of the mutation and the basic characteristics of genes through analyzing the constitution, structure and functional unit. Meanwhile, gene expression will be studied through promoter methylation, RNA transcription and protein translation with the techniques of BSP-PCR, qPCR, WB and IHC. Further, the interaction regulatory networks will be obtained via transcriptome sequencing of cells, ChIP, RIP, Co-IP and RNA Pull Down. The results will provide theoretical foundation for clarifying the function and mechanism of DAZ gene family in the BMI spermatogenesis.