摘要
【目的】基于猪极端饲料报酬表型,利用靶向切割标签技术(cleavage under targets and tagmentation,CUT&Tag)构建H3K27ac差异图谱,鉴定关联的启动子、增强子和关联候选基因,为猪饲料报酬解析提供基础数据。【方法】从209头杜洛克猪中筛选饲料报酬表型差异最大的4头个体分别组成低剩余采食量组(L组,n=2)和高剩余采食量组(H组,n=2)。利用CUT&Tag技术,以H3K27ac作为活性启动子和增强子的表观遗传标记对4头个体的肝脏组织进行全基因组扫描,鉴定关联启动子、增强子和候选基因,并进一步采用real-time quantitative PCR(RT-qPCR)对重要基因进行验证。【结果】共鉴定出47271个H3K27ac峰,包括15739个潜在启动子和31532个推定增强子,大部分的H3K27ac峰位于近端启动子区域。根据H3K27ac全基因的分布情况和作用特点,发现3087个差异峰(对应2188个基因),其中867个H3K27ac峰(对应664个基因)富集在H组,2220个H3K27ac峰(对应1575个基因)富集在L组。GO和KEGG结果发现H3K27ac增强子可能是通过影响胆汁酸代谢、淋巴细胞代谢和NF-κB通路来调节饲料报酬。RT-qPCR结果显示,与H组相比,DIAPH3、DPYD、FTO、TCF7L2、ABCC2、ABCC11、RXRA和ABCG8在L组中显著上调(P<0.05),而PAX5、GRHPR、NFATC1、CARD11、BLNK和LYN在L组中显著下调(P<0.05)。【结论】本研究成功生成了具有极端RFI差异的杜洛克猪肝脏组织中的H3K27ac差异图谱以及相关数据。基于该图谱鉴定和验证了关联候选基因,为进一步解析启动子和增强子调控饲料报酬的分子机制提供了基础数据。
【Objective】Based on the extreme feed reward phenotypes of pigs,the Cleavage Under Targets and Tagmentation(CUT&Tag)technique was used to construct the H3K27ac differential map and identify the associated promoters,enhancers,and candidate genes,thereby providing foundational data for the analysis of feed reward in pigs.【Method】In this study,we screened four individuals with the greatest phenotypic differences in feed compensation from 209 Duroc pigs to form a low residual feed intake group(L,n=2)and a high residual feed intake group(H,n=2),respectively.Liver tissues of the four individuals were scanned genome-wide using CUT&Tag technology with H3K27ac as an epigenetic marker for active promoters and enhancers to identify the associated promoters,enhancers,and candidate genes.Key genes were further validated using Real-time Quantitative PCR(RT-qPCR).【Result】A total of 47271 H3K27ac peaks,including 15739 potential promoters and 31532 putative enhancers,were identified in this experiment,and most of them were located in the proximal promoter region.Based on the distribution and role characteristics of the whole H3K27ac gene,this paper identified 3,087 differential peaks(corresponding to 2188 genes),of which 867 H3K27ac peaks(corresponding to 664 genes)were enriched in the H group,and 2220 H3K27ac peaks(corresponding to 1575 genes)were enriched in the L group.The GO and KEGG results revealed that the H3K27ac enhancers may regulate feed reward by affecting bile acid metabolism,lymphocyte metabolism,and NF-κB pathway.RT-qPCR results showed that DIAPH3,DPYD,FTO,TCF7L2,ABCC2,ABCC11,RXRA,and ABCG8 were significantly up-regulated(P<0.05)in group L compared to group H,while PAX5,GRHPR,NFATC1,CARD11,BLNK and LYN were significantly downregulated in group L(P<0.05).【Conclusion】In this study,we successfully generated an H3K27ac differential map and related data in liver tissues of Duroc pigs with extreme RFI differences.Based on this map,the associated candidate genes were identified and validated,providing basic data for further resolving the molecular mechanism of promoter and enhancer regulation of feed efficiency.
作者
陈栋
王书杰
赵真坚
申琦
余杨
崔晟頔
王俊戈
陈子旸
吴平先
唐国庆
CHEN Dong;WANG Shujie;ZHAO Zhenjian;SHEN Qi;YU Yang;CUI Shengdi;WANG Junge;CHEN Ziyang;WU Pingxian;TANG Guoqing(College of Animal Science and Technology,Sichuan Agricultural University/State Key Laboratory of Swine and Poultry Breeding Industry/Key Laboratory of Livestock and Poultry Multi-omics,Ministry of Agriculture and Rural Affairs/Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province,Chengdu 611130,China;National Center of Technology Innovation for Pigs,Chongqing 402460,China)
出处
《四川农业大学学报》
北大核心
2025年第6期1616-1624,共9页
Journal of Sichuan Agricultural University
基金
国家生猪技术创新中心先导科技项目(NCTIP-XD/B01)
四川省科技厅项目(2020YFN0024、2021ZDZX0008、2021YFYZ0030)
四川省猪创新团队(SCCXTD-2022-08)。
