中国畜牧兽医2025，Vol.52Issue(2)：499-511,13.DOI:10.16431/j.cnki.1671-7236.2025.02.001
灵丘青背山羊ACSL1基因克隆、生物信息学分析及其在脂肪细胞分化过程中的表达研究
Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats
摘要
Abstract
[Objective]This study was aimed to clone the CDS sequence of long-chain acyl-CoA synthetase 1(ACSL1)gene in Lingqiu Qingbei goats and carry out bioinformatics analysis,and explore the expression pattern of ACSL1 gene in different tissues and the differentiation of subcutaneous fat cells in Lingqiu Qingbei goats.[Method]Based on ACSL1 gene sequence of Capra hircus in GenBank(accession No.:XM_018041882.1),specific primers were designed.The CDS sequence of ACSL1 gene was amplified by PCR using the cDNA template of liver in Lingqiu Qingbei goats and cloned and sequenced.The similarity alignment and phylogenetic tree construction were carried out with other species,and the bioinformatics analysis of ACSL1 protein was performed by the online software.The expression of ACSL1 gene in different tissues in Lingqiu Qingbei goats and ACSLA,ACACA,PPARγ and FASN genes at different differentiation stages of subcutaneous fat cells in goats were analyzed by Real-time quantitative PCR.[Result]The CDS region of ACSL1 gene in Lingqiu Qingbei goats was 2 100 bp in length,which encoded 699 amino acids.Similarity alignment revealed that the amino acid sequence of ACSL1 protein in Lingqiu Qingbei goats had the highest similarity with Ovis aries,reaching 99.0％.The results of phylogenetic tree showed that Lingqiu Qingbei goats had the closest relationship with Ovis aries and the farthest relationship with Orcinus orca,and the evolutionary process was highly conservative in different species.Bioinformatics analysis showed that the molecular formula of ACSL1 protein in Lingqiu Qingbei goats was C3529H5567N929O1000S36,the theoretical molecular weight was 78.2 ku,the theoretical isoelectric point was 7.46,the half-life was 30 h,and the instability coefficient was 31.61.ACSL1 protein was an alkaline hydrophobic protein with a transmembrane structure and no signal peptide.ACSL1 protein had 49 phosphorylation sites,and mainly played a biological function in mitochondria.The secondary structure of ACSL1 protein in Lingqiu Qingbei goats was composed of alpha helix(39.63％),random coil(31.33％),extended chain(20.74％)and beta turn(8.30％).ACSL1 protein might interact with 10 proteins such as FASN,ACACA and LPL.The tissue expression analysis showed that ACSL1 gene was expressed in all tissues of Lingqiu Qingbei goats,with the highest expression in liver,which was significantly higher than that in other tissues(P＜0.05),followed by subcutaneous fat,kidney and heart.The results of temporal expression of genes in goat adipocytes at different differentiation stages showed that,compared with day 0 of differentiation,the expression of ACSL1 gene in goats reached the peak on the 8th day of differentiation(P＜0.05),the expression of ACACA and FASN genes reached the peak on the 10th day of differentiation(P＜0.05),and the expression of PPARγ gene reached the peak on the 6th day of differentiation(P＜0.05).[Conclusion]The CDS sequence of ACSL1 gene in Lingqiu Qingbei goats was successfully cloned,and the expression pattern of ACSL1 gene in different tissues and the differentiation of subcutaneous fat cells of goats were clarified.The expression of ACSL1 gene was high in liver and subcutaneous fat.In the process of adipocyte differentiation,the expression of ACSL1,ACACA,PPARγ and FASN genes showed a trend of increasing first and then decreasing.The results provided a theoretical basis for further exploring the molecular mechanism of ACSL1 gene in fat deposition of goats.关键词
灵丘青背山羊/ACSL1基因/克隆/生物信息学/组织表达/脂肪细胞分化Key words
Lingqiu Qingbei goats/ACSL1 gene/cloning/bioinformatics/tissue expression/adipocyte differentiation分类
农业科技引用本文复制引用
艾小楠,程俐芬,白璞,陈正灏,薛丽娜,周胜花..灵丘青背山羊ACSL1基因克隆、生物信息学分析及其在脂肪细胞分化过程中的表达研究[J].中国畜牧兽医,2025,52(2):499-511,13.基金项目
山西省重点研发计划项目(201903D221003) （201903D221003）
山西省现代农业产业技术体系建设专项(2023CYJSTX14) （2023CYJSTX14）
山西省重点研发计划项目(2022ZDYF114) （2022ZDYF114）
山西农业大学科技创新提升工程项目(CXGC2023013) （CXGC2023013）
