基于RNA-Seq的无量山乌骨鸡肝脏组织脂代谢相关基因筛选及其表达分析

doi:10.11843/j.issn.0366-6964.2023.03.012

摘要/Abstract

摘要： 旨在通过分析不同日龄无量山乌骨鸡肝组织mRNA表达谱，挖掘肝脂代谢相关基因及其所参与的调控通路，并确定部分关键基因在肝发育中的表达特征。本研究采集1日龄(D1)和168日龄(D168)各3只母鸡肝组织样品，利用DNBSEQ平台进行转录组测序，随机选取10个差异表达基因进行RT-qPCR验证。以|log₂FC|≥2、Q value≤0.01和差异基因KEGG分析筛选参与脂代谢的差异表达基因，对筛选出的脂代谢相关差异表达基因进行GO富集、KEGG通路和蛋白互作分析。100只雌性雏鸡以出壳体重组间无差异随机分为5组，每组20只鸡，在 D1、D42、D84、D126和D168共5个阶段每组随机屠宰2只健康母鸡(n=10)，并提取肝组织RNA，RT-qPCR构建部分差异基因mRNA表达谱。结果，筛选出50个与脂代谢相关的差异表达基因，其中38个基因表达上调，12个基因表达下调。差异表达基因主要富集在脂质代谢、氧化还原、脂肪酸生物合成、脂肪酸代谢、胆固醇生物合成和甾醇生物合成等过程；差异表达基因参与了类固醇生物合成、脂肪酸代谢、甘油脂类代谢、不饱和脂肪酸生物合成和PPAR信号通路。蛋白互作分析筛选出SCD、ACSBG2、SQLE、HSD17B7、LCAT和LPIN1等关键蛋白基因，通过其mRNA表达量不同参与脂代谢调控。通过转录组测序挖掘出与脂代谢相关的差异表达基因50个，且42个差异表达基因之间存在相互作用关系，该研究结果为进一步解析优质鸡脂代谢相关基因的分子调控提供了理论依据。

关键词: 无量山乌骨鸡, RNA-Seq, 肝组织, 脂代谢, 差异表达基因

Abstract: This study was conducted to screen out and verify differentially expressed genes and their regulatory pathways related to lipid metabolism by analyzing mRNA expression profiles in liver tissues of Wuliangshan Sooty chicken at different ages, and to provide theoretical basis for determining the expression characteristics of genes related to lipid metabolism in liver tissues of Wuliangshan Sooty chicken. In this study, D1 and D168 liver tissues were sequenced by RNA-Seq technique, and there were 3 biological replicates in each stage, transcriptome sequencing using the DNBSEQ platform, 10 differentially expressed genes were randomly selected for RT-qPCR validation. The differential genes related to lipid metabolism were screened by |log₂FC|≥2, Q value≤0.01 and KEGG pathways analysis. The differentially expressed genes of lipid metabolism were analyzed for GO enrichment, KEGG pathways, and protein interactions. A total of 100 female chicks were randomly divided into 5 groups with 20 chickens in each group. Two healthy hens were randomly slaughtered in each group at 5 stages (D1, D42, D84, D126, D168), and RNA was extracted from liver tissues. RT-qPCR was used to construct partial differential genes mRNA expression profiles. A total of 50 differentially expressed genes related to lipid metabolism were screened by transcriptome sequencing, among which 38 genes were up-regulated and 12 genes were down-regulated. The differentially expressed genes were mainly enriched in lipid metabolism, redox process, fatty acid biosynthesis, fatty acid metabolism, cholesterol biosynthesis and sterol biosynthesis. The differentially expressed genes were involved in steroid biosynthesis, fatty acid metabolism, glycerolipids metabolism, unsaturated fatty acid biosynthesis and PPAR signaling pathway. Key protein genes such as SCD, ACSBG2, SQLE, HSD17B7, LCAT and LPIN1 were screened out by protein interaction analysis, which were involved in the regulation of lipid metabolism through their mRNA expression levels. The 50 differentially expressed genes related to lipid metabolism were identified by transcriptome sequencing, and there are interactions among 42 differentially expressed genes. It provides a theoretical basis for further analysis of the molecular regulation of fat metabolism related genes in indigenous Chinese chicken.

Key words: Wuliangshan Sooty chicken, RNA-Seq, liver tissue, lipid metabolism, differentially expressed genes

中图分类号:

S831.2

欧正淼, 周家文, 刘莉莉, 吴芸, 陈粉粉. 基于RNA-Seq的无量山乌骨鸡肝脏组织脂代谢相关基因筛选及其表达分析[J]. 畜牧兽医学报, 2023, 54(3): 976-988.

OU Zhengmiao, ZHOU Jiawen, LIU Lili, WU Yun, CHEN Fenfen. Screening and Expression Analysis of Genes Related to Lipid Metabolism in Liver Tissue of Wuliangshan Sooty Chicken Based on RNA-Seq[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 976-988.

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