金华猪PPARGC1A基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2020.03.006

Abstract

Abstract: The aim of this study was to investigate the genetic characteristics and expression patterns of PPARGC1A gene in Jinhua pigs and Yorkshire pigs.The Jinhua pigs and Yorkshire pigs were used as samples to extract total RNA from the dorsal lipid tissue,and the coding region and real-time quantitative PCR primers were designed according to the sequence of porcine PPARGC1A gene in GenBank (accession No.:NM_213963.2).The porcine GAPDH gene and β-actin protein were used as internal parameters,and the PPARGC1A gene-encoded protein was applied by various bioinformatics methods.Functional analysis was performed and its expression level in dorsal lipid tissue of Jinhua pigs was detected by Real-time quantitative PCR and Western blotting assay.The results showed that the full length of PPARGC1A gene CDS region was 2 361 bp and encoded 786 amino acids.The protein had a molecular size of 90 336.01 u,with serine (Ser) accounting for the highest proportion (13.7%) and tryptophan (Trp) accounting for the lowest proportion (0.8%).Homology alignment results showed that PPARGC1A gene in Jinhua pigs had high homology with goats,cattle and sheep,which were 95.0%,94.9% and 94.9%,respectively,and was highly conserved in species evolution.The PPARGC1A protein of the instability index in Jinhua pigs was 74.88,belonging to unstable hydrophilic protein,and no transmembrane structure.Its secondary structure consists of alpha helix,extended chain,beta turn and random coil,and the proportion of them were 26.59%,5.73%,5.73% and 61.96%,respectively.The PPARGC1A protein homology modeling obtained a series of complex processes such as folding and bending to obtain a three-level structural model.The results of the Real-time quantitative PCR and Western-blotting were consistent,indicating that the expression of PPARGC1A gene in Jinhua pigs with dorsal lipid tissue was significantly lower than that of lean pigs (P<0.05).This study provides a theoretical basis for the discovery of the molecular biological function of PPARGC1A gene in pig fat deposition.

Key words: Jinhua pigs; PPARGC1A gene; bioinformatics analysis; tissue expression; fat deposition

CLC Number:

S828.9

LIU Yufang, GUO Siwu, ZHANG Qingyang, ZHANG Xin, YANG Junqi, BAI Ying. Cloning,Bioinformatics Prediction and Tissue Expression Analysis of PPARGC1A Gene in Jinhua Pigs[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(3): 696-705.

References

[1] PUIGSERVER P,WU Z,PARK C W,et al.A Cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis[J].Cell,1998,92(6):829-839.
[2] 苏燕,彭姝彬,李智琼,等.PPARGC1A基因Thr394Thr/Gly482Ser多态性与2型糖尿病的关联研究[J].遗传,2008,30(3):304-308. SU Y,PENG S B,LI Z Q,et al.Association study between PPARGC1A Thr394Thr/Gly482Ser polymorphisms and type 2 diabetes[J].Hereditas,2008,30(3):304-308.(in Chinese)
[3] LI J D,FENG Q C,QI Y,et al.PPARGC1A is upregulated and facilitates lung cancer metastasis[J].Experimental Cell Research,2017,359(2):356-360.
[4] ZHANG S,JIANG J,CHEN Z,et al.Relationship of PPARG,PPARGC1A,and PPARGC1B polymorphisms with susceptibility to hepatocellular carcinoma in an eastern Chinese Han population[J].OncoTargets and Therapy,2018,11:4651-4660.
[5] SOYAL S M,ZARA G,FERGER B,et al.The PPARGC1A locus and CNS-specific PGC-1α isoforms are associated with Parkinson's disease[J].Neurobiology of Disease,2019,121:34-46.
[6] 王杰,原清会,张明,等.中国荷斯坦奶牛PPARGC1A基因内含子9的SNP与产奶性状的相关分析[J].四川农业大学学报,2010,28(2):219-222. WANG J,YUAN Q H,ZHANG M,et al.Association analysis between SNP of intron 9 in PPARGC1A gene and milking traits in Chinese Holstein breed[J].Journal of Sichuan Agricultural University,2010,28(2):219-222.(in Chinese)
[7] 刘婷婷,孟军,王建文,等.三个品种马PPARGC1A基因PCR-SSCP多态性分析[J].新疆农业科学,2015,52(10):1926-1931. LIU T T,MENG J,WANG J W,et al.Polymorphism analysis of three different horse types PPARGC1A gene by PCR-SSCP[J].Xinjiang Agricultural Sciences,2015,52(10):1926-1931.(in Chinese)
[8] 安世钰,张国敏,尤佩华,等.PPARGC1A/NRF1/TFAM通路核心基因在湖羊睾丸发育过程中的表达变化[J].南京农业大学学报,2019,42(2):345-351. AN S J,ZHANG G M,YOU P H,et al.Research on the change of PPARGC1A/NRF1/TFAM pathway genes expression in the development of Hu sheep testis[J].Journal of Nanjing Agricultural University,2019,42(2):345-351.(in Chinese)
[9] 韩雪蕾,刘榜,彭勇波,等.猪PPARGC1A基因的遗传变异检测及与胴体性状的关联分析[J].华中农业大学学报,2009,28(5):587-590. HAN X L,LIU B,PENG Y B,et al.Hereditary variation of pig PPARGC1A gene and its association with carcass traits of porcine[J].Journal of Huazhong Agricultural University,2009,28(5):587-590.(in Chinese)
[10] GANDOLFI G,CINAR M U,PONSUKSILI S,et al.Association of PPARGC1A and CAPNS1 gene polymorphisms and expression with meat quality traits in pigs[J].Meat Science,2011,89(4):478-485.
[11] 徐芹芹,刘玉芳,康晓龙,等.滩羊EphA3基因克隆表达分析及生物信息学初步研究[J].中国畜牧杂志,2019,3:33-38. XU Q Q,LIU Y F,KANG X L,et al.Cloning,expression and bioinformatics analysis of EphA3 gene in Chinese Tan sheep[J].Chinese Journal of Animal Science,2019,3:33-38.(in Chinese)
[12] 瞿秋红,黄叶,蒋钦杨,等.影响猪脂肪沉积主要候选基因的研究进展[J].动物营养学报,2019,7:2981-2987. QU Q H,HUANG Y,JIANG Q Y,et al.Research progress of major candidate genes affecting pig fat deposition[J].Chinese Journal of Animal Nutrition,2019,7:2981-2987.(in Chinese)
[13] 吕亚宁,叶文文,兰旅涛.牛肌内脂肪沉积影响因素及相关基因研究进展[J].中国草食动物科学,2019,4:55-57. LYU Y N,YE W W,LAN L T.Research progress on influencing factors and related genes of fat deposition in bovine muscle[J].China Herbivore Science,2019,4:55-57.(in Chinese)
[14] 农秋雲,刘嘉琪,单体中.猪肉中脂肪酸组成的品种差异及脂肪酸沉积的调控机制[J].动物营养学报,2019,6:2507-2514. NONG Q Y,LIU J Q,SHAN T Z.Breed difference of fatty acid composition and regulation mechanism of fatty acid deposition in pork[J].Chinese Journal of Animal Nutrition,2019,6:2507-2514.(in Chinese)
[15] 张雨,施力光,荀文娟,等.能量水平和激素对脂肪代谢酶基因的调控研究进展[J].动物营养学报,2019,8:3505-3510. ZHANG Y,SHI L G,XUN W J,et al.Research progress of regulation of fat metabolism enzyme gene by energy level and hormone[J].Chinese Journal of Animal Nutrition,2019,8:3505-3510.(in Chinese)
[16] RAMOSLOPEZ O,RIEZUBOJ J I,MILAGRO F I.Association of the Gly482Ser PPARGC1A gene variant with different cholesterol outcomes in response to two energy-restricted diets in subjects with excessive weight[J].Nutrition,2018,47:83-89.
[17] PARDO R,ENGUIX N,LASHERAS J,et al.Rosiglitazone-induced mitochondrial biogenesis in white adipose tissue is independent of peroxisome proliferator-activated receptor γ coactivator-1α[J].PLoS ONE,2011,6(11):e26989.
[18] PEI X,LIU L,CAI J,et al.Haplotype-based interaction of the PPARGC1A and UCP1 genes is associated with impaired fasting glucose or type 2 diabetes mellitus[J].Medicine,2017,96(23):e6941.
[19] 乌日古木拉.蒙古马运动候选基因PPARGC1A多态性和表达量研究[D].呼和浩特:内蒙古农业大学,2012. WURI G.The investigation of polymorphism and expression's quantity on athletics candidate gene PPARGC1A in Mongolian horses[D].Hohhot:Inner Mongolia Agricultural University,2012.(in Chinese)
[20] 梁爽,王伟伟,孙力,等.PPARGC1启动子甲基化与Ⅱ型糖尿病发病相关性分析[J].分子诊断与治疗杂志,2019,2:91-95. LIANG S,WANG W W,SUN L,et al.Correlation analysis between PPARGC1 promoter methylation and type Ⅱ diabetes[J].Journal of Molecular Diagnosis and Therapy,2019,2:91-95.(in Chinese)
[21] WANG B,NIE W,FU X.Neonatal vitamin A injection promotes cattle muscle growth and increases oxidative muscle fibers[J].Journal of Animal Science and Biotechnology,2019,1:140-149.
[22] 李密杰.秦川牛PPARGC1A基因功能验证及遗传变异研究[D].杨凌:西北农林科技大学,2013. LI M J.Function analysis and genetic variations of PPARGC1A gene in Qinchuan cattle[D].Yangling:Northwest A&F University,2013.(in Chinese)
[23] POPOV D V,LYSENKO E A,MAKHNOVSKII P A,et al.Regulation of PPARGC1A gene expression in trained and untrained human skeletal muscle[J].Physiological Reports,2017,5(23):doi:10.14814/phy2.13543.
[24] AUDREY C,AUDREN F,ALESSIA B,et al.Hepatocyte nuclear factor-1β controls mitochondrial respiration in renal tubular cells[J].Journal of the American Society of Nephrology,2017,28(11):3205-3217.
[25] VOILLET V,CRISTOBAL M S,PERE M C,et al.Integrated analysis of proteomic and transcriptomic data highlights late fetal muscle maturation process[J].Molecular & Cellular Proteomics,2018,17(4):672-693.

Cloning,Bioinformatics Prediction and Tissue Expression Analysis of PPARGC1A Gene in Jinhua Pigs

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[2]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[3]	TAO Qinghua, HUANG Caiyun, CHEN Chao, LIU Wei, SUN Ningning, LI Ang. Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1296-1306.
[4]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[5]	LI Mengyuan, LIU Mengqian, KANG Jiajia, HUANG Ying, DUAN Yong, JIA Junjing, GE Changrong, CAO Weina. Effects of Guanidinoacetic Acid on Meat Quality and Abdominal Fat Deposition in Broilers [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1385-1394.
[6]	HU Leyu, HU Xinyao, LI Ying, ZHAO Shengmiao, SHEN Fengzhen, WANG Changfa, LI Liangliang, WANG Tongtong, REN Huiying. Bioinformatics Analysis of HO-1 Gene and Preparation of Polyclonal Antibody in Dezhou Donkey [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1499-1510.
[7]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[8]	LI Dan, ZHANG Haisen, WANG Yiqun, GAO Dengke, ZHAO Hongcong, LI Chao, JIN Yaping, CHEN Huatao. Eukaryotic Expression Vector Construction,Bioinformatics and Tissue Expression Profiles Analysis of CRY1 Gene in Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 870-881.
[9]	ZHANG Yongjie, SUI Zhiyuan, ZHANG Zhishuai, WANG Chenguang, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1048-1058.
[10]	WEI Yirong, ZHENG Zihua, ZHANG Sanbao, SONG Ying, JIANG Haiyu, SUN Wenyue, CHENG Pengfei, LIU Yufan, ZOU Jianwei, HUANG Yanna, PAN Yan, JIANG Qinyang. Cloning,Bioinformatics Analysis and Tissue Expression of UCP1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 440-450.
[11]	CHEN Chuanhe, LIU Jiali, ZHANG Lilan, ZHAO Ying, TAO Cong. Bioinformatics Analysis of Porcine SGK Family Genes and Their Expression in Porcine Adipose Tissues and Adipocytes [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3310-3320.
[12]	TANG Jingyu, DU Hanyu, MENG Chunchun, LIU Guangqing. Cloning,Bioinformatics Analysis and Subcellular Localization of Interferon-stimulated Gene 15 in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2843-2854.
[13]	NIE Jingru, MA Li, YAN Dawei, DENG Jun, ZHANG Hao, ZHANG Bo, LIU Jinqiao, DONG Xinxing. Analysis of Differentially Expressed Genes and Regulation Pathways of Intramuscular Fat Deposition in Large Diqing Tibetan Pigs at Different Growth Stages [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2855-2868.
[14]	FU Yongzhi, MA Xiaojun, WANG Maohan, LI Zhe, CHEN Zhiyuan, QUAN Wei, ZHAO Han, SU Yongmei, WU Jianhua, ZHU Wenjin. Bioinformatics Analysis and Tissue Expression of MSTN Gene in Yangyuan Donkeys [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2910-2919.
[15]	ZOU Jianwei, ZOU Juhong, SHEN Yujian, WEI Yi, ZHANG Sanbao, LIAN Zitong, XU Jianjian, SONG Ying, HUANG Yanna, WEI Yingming, JIANG Qinyang, ZHENG Zihua. Bioinformatics Analysis, Construction of Eukaryotic Expression Vector and Expression of LMCD1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2033-2042.