黔北麻羊UCP2基因组织表达及其多态性与生长性状的关联分析

doi:10.16431/j.cnki.1671-7236.2020.07.014

摘要/Abstract

摘要： 本研究旨在探讨黔北麻羊解耦联蛋白2（uncoupling protein 2，UCP2）基因的遗传多态性及其与生长性状的关联性，以期为黔北麻羊的育种改良提供理论依据。以200头6月龄黔北麻羊为研究对象，采用直接测序法检测黔北麻羊UCP2基因的SNP位点，并将其与黔北麻羊生长性状进行关联分析；实时荧光定量PCR检测UCP2基因在黔北麻羊不同组织中的表达水平。结果显示，UCP2基因存在4个SNPs位点，分别为：g.29614172 A>G、g.29619320 G>T、g.29619721 C>T和g.29620037 A>G，均产生3种基因型，其中g.29619320 G>T和g.29620037 A>G为错义突变，g.29619320 G>T突变导致甘氨酸（Gly）变为半胱氨酸（Cys），g.29620037 A>G突变导致苏氨酸（Thr）突变为丙氨酸（Ala）。多态信息含量显示，4个SNPs位点均表现为中度多态（0.25 < PIC < 0.5）。χ²检验结果表明，g.29614172 A>G、g.29619721 C>T和g.29620037 A>G突变位点在黔北麻羊群体中均处于Hardy-Weinberg平衡状态（P>0.05），g.29619320 G>T突变位点极显著偏离了Hardy-Weinberg平衡状态（P<0.01）。连锁不平衡分析显示，g.29619320 G>T和g.29619721 C>T位点具有强连锁效应。实时荧光定量PCR结果表明，UCP2基因在黔北麻羊各组织中均有不同程度的表达，其在肺脏中的表达量最高，其次是脾脏、肝脏和心脏，其余组织表达量较低。关联性分析结果表明，g.29614172 A>G位点AG和GG基因型个体体重、体斜长、胸宽、胸深、胸围及管围均显著高于AA基因型，g.29619320 G>T位点TT基因型个体胸深、胸围及管围均显著高于GT和GG基因型，g.29619721 C>T位点CT基因型个体体高显著高于CC和TT基因型，g.29620037 A>G位点GG基因型个体体重、胸深及管围均显著高于AG和AA基因型个体（P<0.05）。本试验结果初步揭示UCP2基因对黔北麻羊部分生长性状有显著影响，发现的4个SNPs位点可作为黔北麻羊经济性状选择的遗传标记。

关键词: 黔北麻羊; UCP2基因; 生长性状; 组织表达

Abstract: This study was aimed to investigate the genetic polymorphism of uncoupling protein 2 (UCP2) gene and its correlation with growth traits in Qianbei Ma goats,in order to provide theoretical basis for breed improvement of Qianbei Ma goats.This experiment took 200 Qianbei Ma goats aged 6 months of age as the research object,SNP of UCP2 gene in Qianbei Ma goats was detected by direct sequencing,and the correlation between UCP2 gene and the growth traits of Qianbei Ma goats was analyzed.The expression of UCP2 gene in different tissues of Qianbei Ma goats was detected by Real-time quantitative PCR.The results showed that there were 4 SNPs in UCP2 gene,which were g.29614172 A>G,g.29619320 G>T,g.29619721 C>T and g.29620037 A>G,all of which produced 3 genotypes.Among them,g.29619320 G>T and g.29620037 A>G were missense mutations,g.29619320 G>T mutation caused glycine (Gly) to cysteine (Cys),and g.29620037 A>G mutation led to threonine (Thr) was mutated to alanine (Ala).The polymorphic information content showed that 4 SNPs were all moderate polymorphism (0.25 < PIC < 0.5).χ² test results showed that g.29614172 A>G,g.29619721 C>T and g.29620037 A>G mutation sites were in Hardy-Weinberg equilibrium in Qianbei Ma goats population (P>0.05),g.29619320 G>T mutation site extremely significantly deviated from Hardy-Weinberg equilibrium (P<0.01).The linkage disequilibrium analysis showed that g.29619320 G>T and g.29619721 C>T sites had strong linkage effects.Real-time quantitative PCR results showed that UCP2 gene was expressed to varying degrees in all tissues of Qianbei Ma goats.The highest expression level was in lung,followed by spleen,liver and heart,and the other tissues had lower expression level.The results of association analysis showed that the body weight,body oblique length,chest width,chest depth,bust and tube circumference of AG and GG genotypes at g.29614172 A>G were significantly higher than that of AA genotype,the chest depth,bust and tube circumference of TT genotype at g.29619320 G>T were significantly higher than that of GT and GG genotypes,the body height of CT genotype at 29619721 C>T was significantly higher than that of CC and TT genotypes,the body weight,chest depth and tube circumference of GG genotype at g.29620037 A>G were significantly higher than that of AG and AA genotypes (P<0.05).The results initially revealed that UCP2 gene had significant effects on some growth traits of Qianbei Ma goats,and 4 SNPs could be used as genetic markers for economic trait selection of Qianbei Ma goats.

Key words: Qianbei Ma goats; UCP2 gene; growth traits; tissue expression

中图分类号:

S813.3

艾锦新, 龙安炬, 罗卫星, 蔡惠芬. 黔北麻羊UCP2基因组织表达及其多态性与生长性状的关联分析[J]. 中国畜牧兽医, 2020, 47(7): 2093-2103.

AI Jinxin, LONG Anju, LUO Weixing, CAI Huifen. Tissue Expression of UCP2 Gene and Its Association Analysis Between Polymorphism and Growth Traits in Qianbei Ma Goats[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2093-2103.

参考文献

[1] KRAUSS S,ZHANG C Y,LOWELL B B.The mitochondrial uncoupling-protein homologues[J].Nature reviews Molecular Cell Biology,2005,6(3):248-261.
[2] RICQUIER D,BOUILLAUD F.Mitochondrial uncoupling proteins:From mitochondria to the regulation of energy balance[J].The Journal of Physiology,2000,529(1):3-10.
[3] 张雷,文生萍,谷大海,等.UCPs基因多态性与脂肪代谢的相关性研究[J].中国畜牧兽医,2010,37(8):78-83. ZHANG L,WEN S P,GU D H,et al.The polymorphisms of UCPs genes associated with fat metabolism[J].China Animal Husbandry & Veterinary Medicine,2010,37(8):78-83.(in Chinese)
[4] TU N,CHEN H,WINNIKES U,et al.Structural organization and mutational analysis of the human uncoupling protein-2(hUCP2) gene[J].Life Sciences,1999,64(3):41-50.
[5] 赵文明,吴晓敏,张康宁,等.雪山鸡UCP基因编码区SNPs与肌内脂肪酸含量关联分析[J].中国家禽,2012,34(13):27-30. ZHAO W M,WU X M,ZHANG K N,et al.SNPs of UCP gene and its association with intramuscular fatty content in Xueshan chickens[J].China Poultry,2012,34(13):27-30.(in Chinese)
[6] 程甲,程曦,朱庆鸿,等.鸡解耦联蛋白基因型与活性氧含量、初生重的相关性分析[J].农业生物技术学报,2011,19(1):102-106. CHENG J,CHENG X,ZHU Q H,et al.Associated analysis on polymorphisms of UCP genotypes,reactive oxygen species contents and birth weight with three chicken breeds[J].Journal of Agricultural Biotechnology,2011,19(1):102-106.(in Chinese)
[7] 郝军,柴志欣,王吉坤,等.麦洼牦牛UCP基因多态性及其与生长性状的遗传效应分析[J].黑龙江畜牧兽医,2019,11:149-153. HAO J,CHAI Z X,WANG J K,et al.Analysis of UCP gene polymorphism and its genetic effects associated with growth traits in Maiwa yak[J].Heilongjiang Animal Science and Veterinary Medicine,2019,11:149-153.(in Chinese)
[8] 国鹏,封元,顾亚玲,等.黑安格斯牛UCP基因多态性位点与生长性状的关联分析[J].黑龙江畜牧兽医,2019,8:6-9. GUO P,FENG Y,GU Y L,et al.Association analysis of UCP gene polymorphism loci and growth traits in Black Angus cattle[J].Heilongjiang Animal Science and Veterinary Medicine,2019,8:6-9.(in Chinese)
[9] MOSTYN A,LITTEN J C,PERKINS K S,et al.Influence of size at birth on the endocrine profiles and expression of uncoupling proteins in subcutaneous adipose tissue,lung,and muscle of neonatal pigs[J].American Journal of Physiology Regulatory Integrative & Comparative Physiology,2005,288(6):1536-1542.
[10] DE OLIVEIRA B A P,DE SOUZA PINHEL M A,NICOLETTI C F,et al.UCP2 and PLIN1 expression affects the resting metabolic rate and weight loss on obese patients[J].Obesity Surgery,2016,27(2):343-348.
[11] 刘建华.绵羊UCP2和UCP3基因的克隆、表达及其遗传变异研究[D].太谷:山西农业大学,2013. LIU J H.Cloning,expression and genetic variation of UCP2 and UCP3 genes in sheep[D].Taigu:Shanxi Agricultural University,2013.(in Chinese)
[12] 师涛,闫晓茹,潘洋洋,等.miR-132-3p靶向UCP2调节绵羊前体脂肪细胞分化的研究[J].畜牧兽医学报,2017,48(11):2059-2067. SHI T,YAN X R,PAN Y Y,et al.Study on the regulation of miR-132-3p in ovine preadipocyte differentiation by targeting UCP2[J].Chinese Journal of Animal and Veterinary Sciences,2017,48(11):2059-2067.(in Chinese)
[13] RICQUIER D,BOUILLAUD F.The mitochondrial uncoupling protein:Structural and genetic studies[J].Progress in Nucleic Acid Research and Molecular Biology,1997,56:83-108.
[14] 李艳华,郦汉杰,赵兴波,等.猪UCP2基因5'调控区和外显子1的遗传变异研究[J].生物化学与生物物理进展,2006,33(3):262-266. LI Y H,HAN H J,ZHAO X B,et al.Research on genetic variation of 5'regulatory region and exon1 of UCP2 in pigs[J].Progress in Biochemistry and Biophysics,2006,33(3):262-266.(in Chinese)
[15] 罗卫星,史忠辉,刘贞德,等.黔北麻羊种质特性研究初报[J].山地农业生物学报,2010,29(5):466-470. LUO W X,SHI Z H,LIU Z D,et al.The preliminary investigation in characteristics of Qianbei Ma goat[J].Journal of Mountain Agriculture and Biology,2010,29(5):466-470.(in Chinese)
[16] 罗卫星,张启林,穆林,等.黔北麻羊的肉用性能和肉质特性研究[J].西南农业学报,2010,23(5):1706-1710. LUO W X,ZHANG Q L,MU L,et al.Studies on mutton performance and quality of Qianbei Ma goat[J].Southwest China Journal of Agricultural Sciences,2010,23(5):1706-1710.(in Chinese)
[17] 徐怀超,昝林森,王洪宝,等.CRTC3基因多态性及基因型组合与秦川牛生长性状的关联分析[J].畜牧兽医学报,2016,47(11):2184-2190. XU H C,ZAN L S,WANG H B,et al.Association of CRTC3 gene polymorphisms and genotype combination with growth traits of Qinchuan cattle[J].Chinese Journal of Animal and Veterinary Sciences,2016,47(11):2184-2190.(in Chinese)
[18] WU X L,MACNEIL M D,DE S,et al.Evaluation of candidate gene effects for beef backfat via Bayesian model selection[J].Genetica (Dordrecht),2005,125(1):103-113.
[19] BRAND M D,ESTEVES T C.Physiological functions of the mitochondrial uncoupling proteins UCP2 and UCP3[J].Cell Metabolism,2005,2(2):85-93.
[20] 蒋秋斐,罗晓瑜,洪龙,等.宁夏西门塔尔杂种牛UCP2基因多态性及其与生长性状的关联性分析[J].中国畜牧兽医,2016,43(7):1843-1849. JIANG Q F,LUO X Y,HONG L,et al.Correlation analysis between UCP2 gene and growth traits in Ningxia Hibrid Simmental cattle[J].China Animal Husbandry & Veterinary Medicine,2016,43(7):1843-1849.(in Chinese)
[21] 高爱琴,李金泉,李宁,等.绵羊FGF5基因SNP的生物信息学分析[J].中国畜牧杂志,2008,44(5):5-7. GAO A Q,LI J Q,LI N,et al.Bioinformatics analysis of sheep FGF5 gene SNP[J].Chinese Journal of Animal Science,2008,44(5):5-7.(in Chinese)
[22] 马晓萌,轩俊丽,王慧华,等.TXNRD1基因多态性与乌珠穆沁绵羊生长性状的关联分析[J].畜牧兽医学报,2016,47(5):909-921. MA X M,X J L,WANG H H,et al.TXNRD1 gene polymorphisms and its association with growth traits in Ujumqin sheep[J].Chinese Journal of Animal and Veterinary Sciences,2016,47(5):909-921.(in Chinese)
[23] 赵聪哲,罗晓彤,李兆华,等.大白猪MC4R、CDC16基因多态性及其与生长性状的关联分析[J].中国畜牧兽医,2019,46(3):792-799. ZHAO C Z,LUO X T,LI Z H,et al.Association between polymorphism of MC4R and CDC16 genes and growth traits in Large White pigs[J].China Animal Husbandry & Veterinary Medicine,2019,46(3):792-799.(in Chinese)
[24] SHERMAN E L,NKRUMAH J D,MURDOCH B M,et al.Polymorphisms and haplotypes in the bovine neuropeptide Y,growth hormone receptor,ghrelin,insulin-like growth factor 2,and uncoupling proteins 2 and 3 genes and their associations with measures of growth,performance,feed efficiency,and carcass merit in beef cattle[J].Journal of Animal Science,2008,86(1):1-16.
[25] 陈翠,张立敏,陈晓杰,等.UCP2基因遗传多态性与西门塔尔牛生长相关性状的遗传效应分析[J].畜牧兽医学报,2012,43(5):708-716. CHEN C,ZHANG L M,CHEN X J,et al.Associations of UCP2 gene polymorphisms with growth traits in Simmental[J].Chinese Journal of Animal and Veterinary Sciences,2012,43(5):708-716.(in Chinese)
[26] 孙国权,高树新,吴慧光,等.解偶联蛋白1、2和3基因在中国西门塔尔牛组织器官中的表达水平及其与胴体品质关系分析[J].华北农学报,2014,29(4):116-120. SUN G Q,GAO S X,WU H G,et al.Analyze the UCP1,UCP2,UCP3 genes expression level in tissues/organs and the relationship with the carcass traits in Chinese Simmental cattle[J].Acta Agriculturae Boreali-Sinica,2014,29(4):116-120.(in Chinese)
[27] 吴雪.江口萝卜猪UCP和STAT3基因变异与组织表达规律研究[D].贵阳:贵州大学,2019. WU X.Study on genetic variation and expression of UCP and STAT3 genes in Jiangkou Luobo pigs[D].Guiyang:Guizhou University,2019.(in Chinese)