HMOX1调控无量山乌骨鸡腹部脂肪沉积的作用及机制研究

鸡腹部脂肪沉积的分子调控机制是改善鸡肉质遗传育种的重要科学问题。无量山乌骨鸡是具有“乌皮乌骨乌肉”特点的优良地方品种，申请人前期研究发现血红素加氧酶1（HMOX1）是决定无量山乌骨鸡腹部脂肪代谢的差异基因，但其对鸡脂肪细胞分化及脂代谢的具体作用及机制尚不清楚。因此，本项目拟以无量山乌骨鸡及其前脂肪细胞为材料，检测不同发育阶段乌骨鸡HMOX1的表达、脂质积累及脂成分分析，明确HMOX1表达量与体脂沉积的相关性；采用过表达和CRISPR-Cas9基因敲除技术，分析HMOX1对脂肪细胞脂质积累及脂代谢的调控作用；同时，结合RNA-Seq、Co-IP蛋白谱测定和线粒体活性分析结果，确定HMOX1作用的靶基因，绘制调控网路图，阐明HMOX1基因的功能及调控腹部脂肪沉积的机制。研究结果为揭示高原优质地方鸡种脂肪沉积的分子调控机制奠定理论基础，为HMOX1作为标志基因选育鸡腹部脂肪沉积性状提供科学依据。

The biological functions of candidate genes and their regulating mechanisms for abdominal fat are important scientific questions to study using Chinese indigenous chicken species. As a major member of oxidative stress regulation, HMOX1 plays an important role in the regulation of lipid metabolism. However, the regulatory effect of HMOX1 on lipid metabolism in chicken adipocytes is still unknown. This project will use silky fowl and preadipocyte and focus on lipid metabolism and investigate the following aspects. We will use HMOX1 gene overexpression and the CRISPR-Cas9 knockout technology, from the level of transcription and protein detection, the expression level of analysis the correlation of expression level and HMOX1 expression quantity, combined with RNA-Seq, Co-IP protein spectrum determination and mitochondrial activity analysis results, the possible target genes were identified, the changes in the expression level of target genes and the expression of lipid metabolism genes were detected, and the regulatory network was plotted to clarify the function of HMOX1 gene and the mechanism of regulating abdominal fat deposition. The results of this project will provide the theoretical guidance and the scientific basis for HMOX1 gene as a marker-gene in molecular breeding to decrease abdominal fat deposition to improve the meat quality.