秦川肉牛AdipoR1和AdipoR2基因克隆、生物信息学分析及表达研究

doi:10.16431/j.cnki.1671-7236.2021.01.003

Abstract

Abstract: The purpose of the experiment was to use bioinformatics to clone and analyze of the CDS regions of the AdipoR1 and AdipoR2 genes of the new Qinchuan beef line (hereinafter referred to as "Qinchuan beef cattle"),and to explore its expression in different tissues and muscle cells of Qinchuan beef cattle induced differentiation at different time.Taking Qinchuan beef cattle as the research object,the CDS region sequences of AdipoR1 and AdipoR2 genes were amplified by PCR,and their functional structure was predicted using bioinformatics software.At the same time,the expression level of AdipoR1 and AdipoR2 genes in 15 tissues of beef cattle and different time after induction differentiation was obtained using qRT-PCR and analyzed for differences then.The results showed that the full length of AdipoR1 gene coding sequence of Qinchuan beef cattle was 1 128 bp,encoding 375 amino acids,the protein molecular weight was 42 446.41 u,and the theoretical isoelectric point was 6.70.The secondary structure of AdipoR1 protein was mainly composed of α-helix.The prediction results of the secondary and tertiary structures were consistent.AdipoR1 protein was hydrophilic protein and had no signal peptide site.The full length of the AdipoR2 gene coding sequence was 1 161 bp,encoding 386 amino acids,the protein molecular weight was 43 707.70 u,and the theoretical isoelectric point was 6.27.The subcellular localization results showed that AdipoR1 protein was 60.9% likely to be located on the cell membrane,and AdipoR2 protein was 73.9% likely to be located on the cell membrane.The results of the amino acids phylogenetic tree of different species showed that the amino acid sequences encoded by the AdipoR1 and AdipoR2 genes of Qinchuan beef cattle were closest to Bos indicus and Bos mutus,respectively.qRT-PCR results showed that AdipoR1 and AdipoR2 genes were all expressed in 15 tissues of Qinchuan beef cattle such as heart,liver,muscle,and so on.And the expression level in muscle tissues was the highest,which was significantly higher than other tissues (P<0.01).There were also obvious differences in the timing of muscle cell differentiation.The experiment revealed the differences in the expression of AdipoR1 and AdipoR2 genes in different tissues and muscle cells of Qinchuan beef cattle at different time after induction differentiation,in order to lay a foundation for further exploration of the biological functions and regulatory mechanisms of AdipoR1 and AdipoR2 genes.

Key words: Qinchuan beef cattle; AdipoR1 gene; AdipoR2 gene; gene cloning; tissue expression

CLC Number:

S823.8⁺1

CHEN Xingyi, YANG Xinran, SUN Bing, ZAN Linsen. Cloning,Bioinformatics Analysis and Expression of AdipoR1 and AdipoR2 Genes from Qinchuan Beef Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 22-34.

References

[1] 唐妮,王书瑶,齐锦雯,等.脂联素调控脂质代谢的研究进展[J].畜牧兽医学报,2018,49(12):2550-2557. TANG N,WANG S Y,QI J W,et al.Research progress on adiponectin regulating lipid metabolism[J].Acta Veterinaria et Zootechnica Sinica,2018,49(12):2550-2557.(in Chinese)
[2] RUIZ M,STAHLMAN M,BOREN J,et al.AdipoR1 and AdipoR2 maintain membrane fluidity in most human cell types and independently of adiponectin[J].Journal of Lipid Research,2019,60(5):995-1004.
[3] 白曼,赵佳,展微,等.脂联素及其受体的研究进展及在生殖中的作用[J].中国畜牧兽医,2015,42(7):1830-1835. BAI M,ZHAO J,ZHAN W,et al.Research progress on adiponectin and its receptors and their roles in reproduction[J].China Animal Husbandry & Veterinary Medicine,2015,42(7):1830-1835.(in Chinese)
[4] YAMAUCHI T,IWABU M,OKADA-IWABU M,et al.Adiponectin receptors:A review of their structure,function and how they work[J].Best Practice & Research Clinical Endocrinology & Metabolism,2014,28(1):15-23.
[5] 于洪馗,吴立玲.脂联素受体的研究进展[J].现代生物医学进展,2009,9(2):375-378. YU H K,WU L L.Recent advances on adiponectin receptor[J].Progress in Modern Biomedicine,2009,9(2):375-378.(in Chinese)
[6] 杨峰,秦超彬,卢荣华,等.草鱼AdipoR1-B基因克隆、组织分布及其表达的营养调控[J].水产学报,2016,40(7):993-1005. YANG F,QIN C B,LU R H,et al.Molecular cloning,tissue distribution and nutritional regulation of AdipoR1-B gene in grass carp (Ctenopharyngodon idella)[J].Journal of Fisheries of China,2016,40(7):993-1005.(in Chinese)
[7] 姜藻航,贺丹丹,夏梦圆,等.AdipoR2介导C2C12肌管细胞促进3T3-L1前体脂肪细胞的糖代谢能力[J].南京农业大学学报,2018,41(5):911-916. JIANG Z H,HE D D,XIA M Y,et al.Mediation of AdipoR2 in 3T3-L1 preadipocytes co-cultured with C2C12 myotubes for glucose absorption and consumption[J].Journal of Nanjing Agricultural University,2018,41(5):911-916.(in Chinese)
[8] 崔晓兵,韩意,李丽,等.脂联素受体表达调控的研究进展[J].生理科学进展,2011,42(3):169-174. CUI X B,HAN Y,LI L,et al.Research progress in regulation of adiponectin receptors expression[J].Progress in Physiological Sciences,2011,42(3):169-174.(in Chinese)
[9] FASSHAUER M,KLEIN J,NEUMANN S,et al.Adiponectin gene expression is inhibited by beta-adrenergic stimulation via protein kinase A in 3T3-L1 adipocytes[J].FEBS Letters,2001,507(2):142-146.
[10] DING S T,LIU B H,KO Y H.Cloning and expression of porcine adiponectin and adiponectin receptor 1 and 2 genes in pigs[J].Journal of Animal Science,2004,82(11):3162-3174.
[11] 李春霖,龚燕平,田慧,等.脂联素受体在正常Wistar大鼠各组织的分布和表达[J].解放军医学杂志,2005,8:718-719. LI C L,GONG Y P,TIAN H,et al.Distribution and expression of adiponectin receptors in different tissues of normal Wistar rats[J].Medical Journal of Chinese Peoples Liberation Army,2005,8:718-719.(in Chinese)
[12] 罗宗刚,李祥,李明洲,等.脂联素(AdipoQ)及其受体基因(AdipoR1 and AdipoR2)在猪背最长肌和腰大肌中的表达差异[J].农业生物技术学报,2011,19(3):507-512. LUO Z G,LI X,LI M Z,et al.Expression changes of adiponection (AdipoQ) and its receptor genes (AdipoR1 and AdipoR2) in porcine longissimus dorsi and psoas major muscles[J].Journal of Agricultural Biotechnology,2011,19(3):507-512.(in Chinese)
[13] YAMAUCHI T,KAMON J,ITO Y,et al.Cloning of adiponectin receptors that mediate antidiabetic metabolic effects[J].Nature,2003,423(6941):762-769.
[14] JIANG Q,CHENG X,CUI Y,et al.Resveratrol regulates skeletal muscle fibers switching through the AdipoR1-AMPK-PGC-1alpha pathway[J].Food Function,2019,10(6):3334-3343.
[15] 宁越,米雪,陈星伊,等.SMAD1基因的沉默和过表达及对秦川牛原代成肌细胞生肌的影响[J].中国农业科学,2019,52(10):1818-1829. NING Y,MI X,CHEN X Y,et al.Silencing and overexpressing SMAD family member 1(SMAD1) gene and its effect on myogenesis in primary myoblast of Qinchuan cattle (Bos taurus)[J].Scientia Agri-cultura Sinica,2019,52(10):1818-1829.(in Chinese)
[16] 付石军,郭时金,阚德英,等.牛肉质性状相关基因的研究进展[J].中国畜牧兽医,2013,40(9):158-165. FU S J,GUO S J,KAN D Y,et al.Research progress on meat quality traits candidate genes in cattle[J].China Animal Husbandry & Veterinary Medicine,2013,40(9):158-165.(in Chinese)
[17] 欧阳霞辉,徐红伟,张德荣,等.绵羊脂联素受体2基因生物信息学分析[J].西北民族大学学报(自然科学版),2015,36(4):45-50. OUYANG X H,XU H W,ZHANG D R,et al.Bioinformatics of adiponectin receptor 2 gene in sheep,Ovis aries[J].Journal of Northwest Minzu University (Natural Science),2015,36(4):45-50.(in Chinese)
[18] 陈星伊,昝林森.miR-150调控动物成脂分化研究进展[J].中国牛业科学,2019,45(6):45-51. CHEN X Y,ZAN L S.Advances in research on miR-150 regulation of lipogenesis[J].China Cattle Science,2019,45(6):45-51.(in Chinese)
[19] 黄磊涛,吴霞,赖琦,等.脂联素通过调控骨代谢在骨质疏松症中的研究与进展[J].中国组织工程研究,2018,22(12):1944-1949. HUANG L T,WU X,LAI Q,et al.Research advances in adiponectin preventing osteoporosis by bone metabolism regulation[J].Chinese Journal of Tissue Engineering Research,2018,22(12):1944-1949.(in Chinese)
[20] YAMAUCHI T,KADOWAKI T.Adiponectin receptor as a key player in healthy longevity and obesity-related diseases[J].Cell Metabolism,2013,17(2):185-196.
[21] MOCINO-RODRIGUEZ M D,SANTILLAN-BENITEZ J G,DOZAL-DOMINGUEZ D S,et al.Expression of AdipoR1 and AdipoR2 receptors as leptin-breast cancer regulation mechanisms[J].Disease Markers,2017,2017:4862016.
[22] CHEN Y L,TAO J,ZHAO P J,et al.Adiponectin receptor PAQR-2 signaling senses low temperature to promote C.elegans longevity by regulating autophagy[J].Nature Communications,2019,10(1):2602.
[23] 刘爽,王秋月.APPL1与胰岛素抵抗及糖尿病肾病关系的研究进展[J].医学综述,2016,22(24):4811-4814. LIU S,WANG Q Y.Research progress in the correlation between APPL1 and insulin resistance and diabetic kidney disease[J].Medical Review,2016,22(24):4811-4814.(in Chinese)
[24] 夏成,王哲,张才,等.INS、GLN和LP对体外培养牛脂肪细胞内HSL、LP的mRNA丰度的影响[J].畜牧兽医学报,2006,12:1312-1318. XIA C,WANG Z,ZHANG C,et al.Effects of neuroendocrine factors on mRNA abundance of HSL and LP in neonatal calf adipocytes in vitro[J].Acta Veterinaria et Zootechnica Sinica,2006,12:1312-1318.(in Chinese)
[25] 罗铮,翁吉梅,封竣琪,等.瘦素基因的研究进展[J].中国畜牧兽医,2018,45(9):2524-2530. LUO Z,WENG J M,FENG J Q,et al.Research advances on leptin gene[J].China Animal Husbandry & Veterinary Medicine,2018,45(9):2524-2530.(in Chinese)
[26] 安清明,周辉通,吴震洋,等.绵羊脂联素基因(ADIPOQ)多态性及其与生长及胴体性状关联性分析[J].中国农业科学,2019,52(10):1807-1817. AN Q M,ZHOU H T,WU Z Y,et al.Polymorphisms of ADIPOQ gene and their association with growth and carcass traits in sheep[J].Scientia Agricultura Sinica,2019,52(10):1807-1817.(in Chinese)
[27] CHRISTODOULOU A,IERODIAKONOU D,AWOFALA A A,et al.Variants in ADIPOQ gene are linked to adiponectin levels and lung function in young males independent of obesity[J].PLoS One,2020,15(1):e225662.
[28] ABOU-SAMRA M,SELVAIS C M,DUBUISSON N,et al.Adiponectin and its mimics on skeletal muscle:insulin sensitizers,fat burners,exercise mimickers,muscling pills or everything together?[J].International Journal of Molecular Sciences,2020,21(7):2620.
[29] KESHVARI S,HENSTRIDGE D C,NG C,et al.Muscle-specific overexpression of AdipoR1 or AdipoR2 gives rise to common and discrete local effects whilst AdipoR2 promotes additional systemic effects[J].Scientific Reports,2017,7:41792.

Cloning,Bioinformatics Analysis and Expression of AdipoR1 and AdipoR2 Genes from Qinchuan Beef Cattle

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