转录组数据揭示<i>PLA2G4A</i>调控坝上长尾鸡肌肉组织中肌苷酸的特异性沉积

doi:10.7668/hbnxb.20194925

摘要/Abstract

摘要：

旨在探究PLA2G4A基因对坝上长尾鸡腿肌肌肉组织中肌苷酸(IMP)的调控水平,为进一步探究坝上长尾鸡PLA2G4A基因的生物学功能奠定基础。以选取270日龄的坝上长尾鸡腿为试验材料,将坝上长尾鸡腿肌组织以肌苷酸含量高低进行分组,每组各5个样本,从腿肌组织中提取总RNA,对其进行转录组测序分析,并进行KEGG富集分析GO功能注释和蛋白互作网络分析;利用RT-qPCR技术,分析差异基因在坝上长尾鸡腿肌组织中的mRNA表达水平;从测序结果中筛选参与甘油磷酸酯代谢通路的基因PLA2G4A。KEGG富集分析结果表明,与肌苷酸合成相关的信号通路包括甘油磷酸酯代谢、醚脂代谢、花生四烯酸代谢等。GO功能注释表明,PLA2G4A基因参与11个生物学过程,包括甘油磷酸酯代谢,醚脂代谢、花生四烯酸代谢、亚油酸代谢等。蛋白互作发现,PLA2G4A基因与花生四烯酸代谢和甘油磷酸酯代谢过程中的重要调控酶共享功能。RT-qPCR结果显示,坝上长尾鸡腿肌组织中PLA2G4A基因的表达量在样本中存在显著性差异。综上所述,PLA2G4A基因可能是影响鸡肉IMP沉积的主效基因。

关键词: 坝上长尾鸡, 肌苷酸, 转录组, PLA2G4A基因, mRNA

Abstract:

In order to investigate the regulatory level of PLA2G4A gene on inosine monophosphate(IMP)in the leg muscle tissue of Bashang Long-tailed chicken,and further explore the biological function of PLA2G4A gene in Bashang Long-tailed chicken.The leg muscle tissue of 270-day-old Bashang Long-tailed chicken was categorized based on the inosine monophosphate content,with five samples allocated to each group.Transcriptome sequencing analysis was conducted on the muscle tissue,followed by KEGG enrichment analysis,GO function annotation and protein interaction network analysis.RT-qPCR was used to analyze the mRNA expression level gene in the leg muscle tissue of Bashang Long-tailed chicken.KEGG enrichment analysis showed that the signal pathways related to IMP synthesis included glycerophospholipid metabolism,etherlipid metabolism and arachidonic acid metabolism.GO functional annotation showed that the PLA2G4A gene was involved in 11 biological processes,including glycerophosphate metabolism,etherlipid metabolism,arachidonic acid metabolism and linoleic acid metabolism.Protein interaction revealed that the PLA2G4A gene shared important regulatory enzyme functions in arachidonic acid metabolism and glycerophosphate metabolism.The Real-time Fluorescence Quantitative PCR results showed that there was a significant difference in the expression level of the PLA2G4A gene in the leg muscle tissue of Bashang Long-tailed chicken in the sample.In summary,the PLA2G4A gene may be the main gene affecting IMP deposition in chicken.

Key words: Bashang Long-tailed chicken, Transcriptome, IMP, PLA2G4A, mRNA

何玲钰, 乔贤, 王新越, 李祥龙. 转录组数据揭示PLA2G4A调控坝上长尾鸡肌肉组织中肌苷酸的特异性沉积[J]. 华北农学报, 2024, 39(S1): 329-336. doi: 10.7668/hbnxb.20194925.

HE Lingyu, QIAO Xian, WANG Xinyue, LI Xianglong. Transcriptome Data Reveals the Specific Deposition of Inosine in Muscle Tissue of Bashang Long-tailed Chickens Regulated by PLA2G4A[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(S1): 329-336. doi: 10.7668/hbnxb.20194925.

http://www.hbnxb.net/CN/Y2024/V39/IS1/329

图/表 6

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	Wang K T, Zhou Y S, Huang S Z, Jiang Y, Fu L, Meng Q L, Ma X J, Ding X D. Single nucleotide polymorphism in PLA2G4A gene and association with feed conversion traits in swine[J]. Chinese Journal of Animal Science, 2019, 55(11):52-58.

基因名称 Gene name	登录号 Accession No.	引物序列(5'-3') Primer sequence(5'-3')	扩增长度/bp Amplification length
PLA2G4A	NC_052539.1	F:5'-CACTTCCCGTGACGTACCAG-3' R:5'-TGGATCCAGAAAGACCAGCG -3'	146
β-actin	396526	F:5'-TATTGCTGCGCTCGTTGTTG-3' R:5'-GGGGTACTTCAGGGTCAGGA-3'	199

基因名称 Gene name	登录号 Accession No.	引物序列(5'-3') Primer sequence(5'-3')	扩增长度/bp Amplification length
PLA2G4A	NC_052539.1	F:5'-CACTTCCCGTGACGTACCAG-3' R:5'-TGGATCCAGAAAGACCAGCG -3'	146
β-actin	396526	F:5'-TATTGCTGCGCTCGTTGTTG-3' R:5'-GGGGTACTTCAGGGTCAGGA-3'	199

样本 Sample	原始reads Raw reads	过滤后reads Clean reads	过滤后碱基/Gb Clean bases	错误发现率/% False discovery rate	Q20/%	Q30/%	GC含量/% GC content
A1	50 728 000	48 889 916	7.33	0.03	97.39	93.09	51.85
A2	42 785 508	40 886 724	6.13	0.03	97.28	92.85	51.58
A3	47 588 060	45 744 878	6.86	0.03	97.19	92.63	52.19
A4	52 503 284	50 632 380	7.59	0.03	96.72	91.73	50.37
A5	43 447 046	41 692 442	6.25	0.03	97.44	93.39	52.31
B1	46 702 784	44 980 912	6.75	0.03	97.29	92.8	52.23
B2	41 813 830	40 168 072	6.03	0.03	97.28	92.84	52.61
B3	43 513 932	42 487 038	6.37	0.03	96.88	91.9	52.94
B4	45 702 690	44 466 824	6.67	0.03	97.03	92.29	50.88
B5	46 914 640	45 820 392	6.87	0.03	97.37	93.1	52.39

样本 Sample	原始reads Raw reads	过滤后reads Clean reads	过滤后碱基/Gb Clean bases	错误发现率/% False discovery rate	Q20/%	Q30/%	GC含量/% GC content
A1	50 728 000	48 889 916	7.33	0.03	97.39	93.09	51.85
A2	42 785 508	40 886 724	6.13	0.03	97.28	92.85	51.58
A3	47 588 060	45 744 878	6.86	0.03	97.19	92.63	52.19
A4	52 503 284	50 632 380	7.59	0.03	96.72	91.73	50.37
A5	43 447 046	41 692 442	6.25	0.03	97.44	93.39	52.31
B1	46 702 784	44 980 912	6.75	0.03	97.29	92.8	52.23
B2	41 813 830	40 168 072	6.03	0.03	97.28	92.84	52.61
B3	43 513 932	42 487 038	6.37	0.03	96.88	91.9	52.94
B4	45 702 690	44 466 824	6.67	0.03	97.03	92.29	50.88
B5	46 914 640	45 820 392	6.87	0.03	97.37	93.1	52.39

信号通路 Signal pathway	基因 Genes	表达 Expression	P值 P-value
甘油磷酸脂代谢Glycerophospholipid metabolism	PLA2G4A^*	下调	0.018 8
醚脂代谢Etherlipid metabolism	PLPP1	下调	0.015 4
脂肪酸降解Fattyacid degradation	ACSL1	上调	0.043 4
花生四烯酸代谢Arachidonic acid metabolism	PLA2G2A	下调	0.022 3
戊糖磷酸途径Pentose phosphate pathway	FBP1	下调	0.002 2

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转录组数据揭示PLA2G4A调控坝上长尾鸡肌肉组织中肌苷酸的特异性沉积

Transcriptome Data Reveals the Specific Deposition of Inosine in Muscle Tissue of Bashang Long-tailed Chickens Regulated by PLA2G4A

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转录组数据揭示PLA2G4A调控坝上长尾鸡肌肉组织中肌苷酸的特异性沉积

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