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

doi:10.16431/j.cnki.1671-7236.2025.02.001

Abstract

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 ACSL1,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.

Key words: Lingqiu Qingbei goats; ACSL1 gene; cloning; bioinformatics; tissue expression; adipocyte differentiation

CLC Number:

S827
Q78

AI Xiaonan, CHENG Lifen, BAI Pu, CHEN Zhenghao, XUE Lina, ZHOU Shenghua. Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(2): 499-511.

References

[1] FU L,SHI J,MENG Q,et al.Verification of key target molecules for intramuscular fat deposition and screening of SNP sites in sheep from Small-tail Han sheep breed and its cross with Suffolk[J].International Journal of Molecular Sciences,2024,25(5):2951.
[2] 郭宏宇,赵鹏,李俊,等.灵丘青背山羊肉品质研究[J].中国草食动物科学,2020,40(5):28-32.GUO H Y,ZHAO P，LI J,et al.Research on the quality of Lingqiu Green back goat[J].Chinese Herbivorous Animal Science,2020,40(5):28-32.(in Chinese)
[3] JIN S,LIU W,JIANG H.Intramuscular fat graft results in lower retention but similar quality compared with subcutaneous counterpart[J]. The Journal of Craniofacial Surgery,2022,33(8):2688-2691.
[4] HAN Y,HE X,YUN Y,et al.The characterization of subcutaneous adipose tissue in Sunit sheep at different growth stages:A comprehensive analysis of the morphology,fatty acid profile,and metabolite profile[J].Foods,2024,13(4):544.
[5] YU X,LIU X,LI Y,et al.Enrichment of C17∶0-rich saturated fatty acids from sheep tail fat for adjuvant therapy of non-small-cell lung cancer[J].Food Science and Biotechnology,2024,33(8):1947-1956.
[6] SCHULZE A,HARRIS A L.How cancer metabolism is tuned for proliferation and vulnerable to disruption [J].Nature,2012,491(7424):364-373.
[7] WU Z M,SUN J,LIAO Z,et al.An update on the therapeutic implications of long-chain acyl-coenzyme A synthetases in nervous system diseases[J].Frontiers in Neuroscience,2022,16:1030512.
[8] CAO Y,YU Y S,ZHANG L C,et al.Transcript variants of long-chain acyl-CoA synthase 1 have distinct roles in sheep lipid metabolism[J].Frontiers in Genetics,2022,13:1021103.
[9] KATO A,ITO M,SANAKI T,et al.Acsl1 is essential for skin barrier function through the activation of linoleic acid and biosynthesis of ω-O-acylceramide in mice[J].Biochimica et Biophysica Acta.Molecular and Cell Biology of Lipids,2022,1867(2):159085.
[10] SINGH A B,DONG B,XU Y Y,et al.Identification of a novel function of hepatic long-chain acyl-CoA synthetase-1(ACSL1) in bile acid synthesis and its regulation by bile acid-activated farnesoid X receptor[J].Biochimica et Biophysica Acta.Molecular and Cell Biology of Lipids,2018,1864(3):358-371.
[11] GREVENGOED T J,COOPER D E,YOUNG P A,et al.Loss of long-chain acyl-CoA synthetase isoform 1 impairs cardiac autophagy and mitochondrial structure through mechanistic target of rapamycin complex 1 activation[J].The FASEB Journal,2015,29(11):4641-4653.
[12] ZHAO L,PASCUAL F,BACUDIO L,et al.Defective fatty acid oxidation in mice with muscle-specific acyl-CoA synthetase 1 deficiency increases amino acid use and impairs muscle function[J].Journal of Biological Chemistry,2019,294:8819-8833.
[13] JESUS V H,RAMAYO C Y,PASSOLS M,et al.Global analysis of the association between pig muscle fatty acid composition and gene expression using RNA-Seq[J].Scientific Reports,2023,13(1):535.
[14] ZHAO Z D,RAZA S H A,TIAN H S,et al.Effects of overexpression of acsl1 gene on the synthesis of unsaturated fatty acids in adipocytes of bovine[J].Archives of Biochemistry and Biophysics,2020,695:108648.
[15] 田宏山,苏晓彤,赵志东,等.siRNA沉默ACSL1基因对秦川肉牛脂肪细胞中不饱和脂肪酸合成的影响[J].农业生物技术学报,2020,28(10):1722-1732.TIAN H S,SU X T,ZHAO Z D,et al.Effects of silencing ACSL1 gene by siRNA on the synthesis of unsaturated fatty acids in adipocytes of Qinchuan beef cattle (Bos taurus)[J].Journal of Agricultural Biotechnology,2020,28(10):1722-1732.(in Chinese)
[16] 梁莎莎,庞春英,邓廷贤,等.奶水牛ACSL1基因组织表达分析及其对脂肪酸代谢相关基因表达的影响[J].中国畜牧杂志,2020,56(5):41-45.LIANG S S,PANG C Y,DENG T X.Expression of ACSL1 and its effect on expression involved in fatty acid metabolism in buffalo[J].Chinese Journal of Animal Science,2020,56(5):41-45.(in Chinese)
[17] 曹阳,金海国,于永生,等.ACSL1基因对绵羊脂肪代谢的影响[J].中国兽医学报,2018,38(4):824-832.CAO Y,JIN H G,YU Y S,et al.Effect of ACSL1 gene on lipid metabolism of sheep[J].Chinese Journal of Veterinary Science,2018,38(4):824-832.(in Chinese)
[18] 周胜花,薛丽娜,陈正灏,等.基于转录组测序技术研究饲粮维生素E调控山羊皮下脂肪中脂肪酸代谢的机理[J].动物营养学报,2023,35(5):3266-3279.ZHOU S H,XUE L N,CHEN Z H,et al.Mechanism of dietary vitamin E regulating metabolism of fatty acids in goat subcutaneous fat based on transcriptome sequencing technology[J].Chinese Journal of Animal Nutrition,2023,35(5):3266-3279.(in Chinese)
[19] YANG Z G,ZHU H B,HYANG X P,et al.Molecular characterization,tissue distribution profile,and nutritional regulation of ACSL1 gene family in golden pompano (Trachinotus ovatus)[J].International Journal of Molecular Sciences,2022,23(12):6437.
[20] BAI Y,LI J,WEI Y,et al.Proteome analysis related to unsaturated fatty acid synthesis by interfering with bovine adipocyte ACSl gene[J].Antioxidants,2024,13(6):641.
[21] SHAN B,YAN M,YANG K,et al.miR-218-5p affects subcutaneous adipogenesis by targeting acsl1,a novel candidate for pig fat deposition[J].Genes,2022,3(2):260.
[22] 张清月,王雪,刘树林,等.绒山羊羔羊和成年羊前体脂肪细胞的原代培养及传代方法[J].动物营养学报,2018,30(9):3551-3558.ZHANG Q Y,WANG X,LIU S L,et al.Primary culture and passage method of preadipocytes of cashmere lambs and adult goats[J].Chinese Journal of Animal Nutrition,2018,30(9):3551-3558.(in Chinese)
[23] 杜琛,付绍印,韩志玲,等.绒山羊肌内前体脂肪细胞的基因表达分析[J].畜牧兽医学报,2013,44(10):1532-1538.DU C,FU S Y,HAN Z L,et al.Gene expression anslysis on intranmuscular preadipocytes in the cashmere goats[J].Acta Veterinaria et Zootechnica Sinica,2013,44(10):1532-1538.(in Chinese)
[24] NAN J,LEE J S,LEE S A,et al.An essential role of the N-terminal region of ACSL1 in linking free fatty acids to mitochondrial β-oxidation in C2C12 myotubes[J].Molecules and Cells,2021,44(9):637-646.
[25] LI J,ZHANG W,WU J,et al.CPT1C-mediated fatty acid oxidation facilitates colorectal cancer cell proliferation and metastasis[J].Acta Biochimica et Biophysica Sinica,2023,55(8):1301-1309.
[26] PEI Y,SONG Y,WANG B,et al.Integrated lipidomics and rna sequencing analysis reveal novel changes during 3T3-L1 cell adipogenesis[J].PeerJ,2022,10:13417.
[27] XIONG L,PEI J,WU X,et al.Explaining unsaturated fatty acids (UFAs),especially polyunsaturated fatty acid (PUFA) content in subcutaneous fat of yaks of different sex by differential proteome analysis[J].Genes,2022,13(5):790.
[28] ZHAO W,SU J,WANG Y,et al.Functional importance of palmitoyl protein thioesterase 1(PPT1) expression by Sertoli cells in mediating cholesterol metabolism and maintenance of sperm quality[J].Molecular Reproduction and Development,2019,86(8):984-998.
[29] HUANG Z R,DENG J C,LI Q Y,et al.Protective mechanism of common buckwheat (Fagopyrum esculentum Moench.) against nonalcoholic fatty liver disease associated with dyslipidemia in mice fed a high-fat and high-cholesterol diet[J].Journal of Agricultural and Food Chemistry,2020,68(24):6530-6543.
[30] ZENG L,ZHOU J Z,ZHANG Y W,et al.Paternal cadmium exposure induces glucolipid metabolic reprogramming in offspring mice via PPAR signaling pathway[J].Chemosphere,2023,339:139592.
[31] GREVENGOED T J,KLETT E L，COLEMAN R A,et al.Acyl-CoA metabolism and partitioning[J].Annual Review of Nutrition,2014,34(1):1-30.
[32] 肖可瑞,雷晨怡,熊钟珉.ACSL1促进肝脏脂质积累与炎症发生并参与疾病发展的研究进展[J].西安交通大学学报，2023,44(4):644-653.XIAO K R,LEI C Y,XIONG Z M,et al.Advances in studies on ACSL1’s promoting hepatic lipid accumulation and inflammation and participating in disease development[J].Journal of Xi’an Jiaotong University (Medical Sciences), 2023,44(4):644-653.(in Chinese)
[33] CHENG H L,CHEN S,XU J H，et al.Molecular cloning and nutrient regulation analysis of long chain acyl-CoA synthetase 1 gene in grass carp,Ctenopharyngodon idella L.[J].Biochemistry & Molecular Biology,2017,204:61-68.
[34] TANG Y,ZHOU J,SHING C,et al.Fatty acid activation in carcinogenesis and cancer development:Essential roles of long-chain acyl-CoA synthetases[J].Oncology Letters,2018,16(2):1390-1396.
[35] TIANZUO Z,MARGARETE P,OBERTR E，et al.Overexpressed FATP1,ACSVL4/FATP4 and ACSL1 increase the cellular fatty acid uptake of 3T3-L1 adipocytes but are localized on intracellular membranes[J].PLoS One,2012,7(9):e45087.
[36] ZHAO Z,ABBAS RAZA S H,TIAN H,et al.Effects of overexpression of ACSL1 gene on the synthesis of unsaturated fatty acids in adipocytes of bovine[J].Archives of Biochemistry and Biophysics,2020,695(1):108648.
[37] WU W,ZHANG J,ZHAO C,et al.CTRP6 regulates porcine adipocyte proliferation and differentiation by the AdipoR1/MAPK signaling pathway[J].Journal of Agricultural and Food Chemistry,2017,65(27):5512-5522.
[38] LV Y,CAO Y,GAO Y,et al.Effect of ACSL3 expression levels on preadipocyte differentiation in Chinese Red steppe cattle[J].DNA and Cell Biology,2019,38(9)：945-954.
[39] YU X,FANG X,XIAO H,et al.The effect of acyl-CoA synthetase long-chain family member 5 on triglyceride synthesis in bovine preadipocytes[J].Archives Animal Breeding,2019,62(1):257-264.
[40] LEE J M,CHOI S S,LEE Y H,et al.The E3 ubiquitin ligase TRIM25 regulates adipocyte differentiation via proteasome-mediated degradation of PPARγ[J].Experimental & Molecular Medicine,2018,50(10):1-11.
[41] 徐小春,陈文娟,赵瑞，等.滩羊肌内前体脂肪细胞的分离培养及分化相关基因的表达规律研究[J].家畜生态学报,2020,41(11):35-41.XU X C,CHEN W J,ZHAO R,et al.Culture and induced differentiation of intramnscular preadupocytes and expression of related genes from Tan sheep[J].Journal of Domestic Animal Ecology,2020,41(11):35-41.(in Chinese)

Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles 0

Metrics

Comments

[1]	XIE Beiyiting, WANG Yue, MENG Chunhua, QIAN Yong, ZHANG Jun, ZHANG Jianli, WANG Huili, CAO Shaoxian, LI Yinxia. Cloning and Sequence Analysis of PSMB9 Gene in Hu Sheep and Its Effect on Myoblast Proliferation [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 1-12.
[2]	TIAN Huihui, WANG Bingxin, ZHU Zhaoyan, YU Yange, DING Mengxia, TIAN Yadong, JIANG Ruirui, SUN Guirong, HAN Ruili, KANG Xiangtao, YAN Fengbin, GUO Yujie. Cloning,Bioinformatics Analysis of HSPA8 Gene and Its Expression Pattern in Thymus in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 39-51.
[3]	CHEN Zuoxin, PAN Yanlin, HUANG Yunzhen, LI Linlin, DONG Jiawen, XIANG Yong, XU Zhihong, SUN Minhua, HUANG Shujian, LIAO Ming, ZHANG Junqin. Isolation,Identification and Whole Genome Sequence Analysis of Two Strains of Tambusu Virus from Waterfowl [J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(1): 310-321.
[4]	ZHOU Fangting. Cloning,Molecular Characteristics and Tissue Differential Expression Analysis of KLF15 Gene in Buffalo [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(9): 3715-3725.
[5]	LI Jinnan, WANG Yahui, DENG Tianyu, LIANG Mang, DU Lili, LI Keanning, XUE Qingqing, QIAN Li, GAO Xue, ZHANG Lupei, ZHU Bo, CHEN Yan, WANG Zezhao, LI Junya, GAO Huijiang. Cloning,Bioinformatics Analysis and Tissue Expression Profile of Coding Region of Bovine MED28 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(8): 3225-3236.
[6]	LI Rong, FU Guowen, ZENG Qin, WU Jiao, YUAN Jiarui, LIU Yabo, SILANG Yuzhen, FAN Yueyuan. Analysis of the Differences of Chemokine-binding Protein Gene Between Ovine Contagious Pustular Dermatitis Virus Oral and Visceral Infection Strains and Their Prokaryotic Expression [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(8): 3506-3518.
[7]	ZHANG Xuedi, XU Kai, YIN Shuo, LIU Jing, WANG Zhonghao, QIN Jianhua, WU Pengyuan, WANG Chuanwen. Cloning,Sequence Analysis and Expression Characteristics of Carboxylesterase 2 Gene in Dermanyssus gallinae [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 2973-2983.
[8]	YUAN Liang, HAN Xiaoxi, GONG Yingchao, FAN Xianan, HAO Beili, HAO Shuangshuang, LI Changwen, CHANG Yicong, LI Rui, LIU Fangping. Bioinformatics Analysis and Prokaryotic Expression of SipD Protein of Salmonella Enteritidis in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(7): 2998-3007.
[9]	JIANG Lei, WANG Yuxin, WAN Yuan, ZAN Linsen, WANG Hongbao. Bioinformatics and Expression Characteristics Analysis of MGP Gene in Qinchuan Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(6): 2261-2272.
[10]	JING Kemin, ZHANG Peng, LI Yuqian, TIAN Yuan, DONG Wenjing, CHAI Zhixin, WANG Jikun, ZHONG Jincheng, CAI Xin. Cloning, Bioinformatic and Expression Analysis of SERPINE1 Gene in Longissimus Dorsi Muscle of Yaks with Different Tenderness [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(6): 2308-2318.
[11]	SUN Shiyu, LI Jinlan, XING Jiani, DONG Yaohui, LI Yanyan, WANG Youli, LIN Yaqiu. Cloning,Bioinformatics Analysis and Spatiotemporal Expression of CKMT2 Gene in Jianzhou Big-eared Goats [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(6): 2342-2353.
[12]	HUANG Liqing, DUAN Anqin, ZHENG Haiying, YANG Chunyan, SHANG Jianghua. Sequence Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of SFRP1 Gene in Buffalo [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1807-1818.
[13]	ZOU Juhong, MO Zhihua, LIU Yufan, ZOU Jianwei, LU Jun, HE Haien, WANG Fan, HUANG Yanna, JIANG Qinyang. Cloning,Bioinformatics Analysis and Eukaryotic Expression Vector Construction of Rheb Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1819-1826.
[14]	PAN Shiyu, YIN Na, WANG Jiaxing, WANG Qiangzhou, CHEN Shihao, BAI Hao, CHANG Guobin. Bioinformatic Analysis of Chicken DCAF1 and Its Impact on the Replication of ALV-J [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1837-1845.
[15]	SUN Can, WEI Shuo, YAN Yujing, ZHAO Tianyi, ZHU Dexin, DENG Xingmei, GUO Jia, ZHANG Hui, SUN Zhihua. Bioinformatics Analysis of Brucella BspF Protein and Its Effect on Expression of Inflammatory Factors in Macrophages [J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1846-1856.