黔北黑猪肌内脂肪沉积相关基因的筛选  

Screening of genes related to intramuscular fat deposition in Qianbei black pigs

作　　者：孟金柱 吴震洋[1] 安清明 赵园园 MENG Jinzhu;WU Zhenyang;AN Qingming;ZHAO Yuanyuan(Guizhou Provincial Key Laboratory for Biodiversity Conservation and Utilization in the Fanjing Mountain Region,Tongren University,Tongren 554300,Guizhou,China;College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]铜仁学院贵州省梵净山地区生物多样性保护与利用重点实验室,贵州铜仁554300 [2]湖南农业大学动物医学院,长沙410128

出　　处：《浙江大学学报（农业与生命科学版）》2021年第6期797-804,共8页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：贵州省教育厅青年科技人才成长项目(黔教合KY字〔2020〕166);贵州省科技计划(黔科合基础〔2020〕1Y138);贵州省重点实验室项目(黔科合平台人才〔2020〕2003号);贵州省铜仁市科学技术局科技支撑计划(铜市科研〔2020〕128号);铜仁学院生态畜牧创新团队项目(CXTD[2020-19])。

摘　　要：选取5头12月龄的健康黔北黑猪,屠宰后分别取其背最长肌(longissimus dorsi muscle,LDM)和腰大肌(psoas major muscle,PMM),一部分用于检测总脂肪含量,另一部分用于提取总RNA、建库及测序。将获得的重叠群与猪RefSeq数据库进行比对,获得注释基因。利用DESeq21.26.0软件分析差异表达的mRNA,并经GOseq 1.15.5软件和KOBAS 3.0软件对差异表达的mRNA进行基因本体(gene ontology,GO)以及京都基因和基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)信号通路分析;然后通过实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)对筛选出的与脂肪沉积相关的候选基因进行验证。结果表明,PMM中的脂肪含量极显著高于LDM(P<0.01)。通过高通量测序,共获得162个差异表达基因,其中66个上调,96个下调。通过GO、KEGG分析并结合功能查找,筛选出6个与脂肪沉积相关的差异表达基因,qRTPCR验证结果显示,LPL、PPARA、COX2、PRKAG2和SCD在LDM和PMM中的表达情况存在显著差异。本研究结果为探究黔北黑猪肌内脂肪沉积的分子机制奠定了基础,也为黔北黑猪的保护、开发和利用提供了实验依据。This study was designed to screen out the key genes related to intramuscular fat deposition through detecting the fat contents in longissimus dorsi muscle(LDM)and psoas major muscle(PMM),and explore the differentially expressed genes between the two types of muscle of Qianbei black pigs using high-throughput sequencing technology.Five 12-month-old healthy Qianbei black pigs were slaughtered,and LDM and PMM were collected.Some of them were used to detect total fat content,and the rest were used for total RNA extraction,library constructing and sequencing.The annotation genes were obtained by comparing the obtained unigenes with RefSeq database of pig.The differentially expressed mRNAs were analyzed by DESeq21.26.0 software,and moreover gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis of differentially expressed mRNAs were analyzed by GOseq 1.15.5 software and KOBAS 3.0 software.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to verify the candidate genes related to fat deposition.The results showed that the fat content in the PMM was significantly higher than that in the LDM(P<0.01).A total of 162 differentially expressed genes were obtained by high-throughput sequencing,of which 66 were up-regulated and 96 were down-regulated.Six differentially expressed genes associated with fat deposition were screened out by GO,KEGG analysis combined with the function searching.qRT-PCR results showed that the expression of LPL,PPARA,COX2,PRKAG2 and SCD existed in significant differences between LDM and PMM.The above results lay a foundation for exploring the molecular mechanism of intramuscular fat deposition in Qianbei black pigs,and further provide an experimental basis for the protection,development and utilization of Qianbei black pigs.

关 键 词：黔北黑猪 背最长肌 腰大肌 差异表达基因 脂肪沉积 

分 类 号：S828.8[农业科学—畜牧学]