为获得中国西门塔尔牛解偶联蛋白（Uncoupling proteins，UCPs） ucp1、ucp2、ucp3 基因的组织表达谱，揭示该基因与能量代谢的关系，探索调控肉牛脂肪沉积的关键基因表达规律，筛选影响肉牛肥育性状的候选基因。以20头中国西门塔尔牛为试验材料，提取其15 种组织（肝脏、淋巴、肋间肉、肩肉、脾脏、肾脏、背最长肌、肺、心脏、肠脂、网脂、睾丸、小肠、胰脏、背膘）总RNA，设计合成 ucp1、ucp2、ucp3 基因的引物，以牛 β-actin 基因为内参，利用实时荧光定量PCR 技术，检测 ucp1、ucp2、ucp3 基因的 mRNA 在以上各组织中的表达。结果表明，ucp1、ucp2、ucp3 基因在睾丸、小肠、心脏、胰腺等组织、器官中均有不同程度的表达，ucp1 在背膘中高表达，胰脏、小肠、睾丸、网脂、肠脂、心脏中度表达，在其他组织中低表达；ucp2 在睾丸中高表达，肺脏、肝脏、背膘中度表达，在其他组织中低表达；ucp 3在背最长肌、肋间肉、肩肉中高表达，肺脏中度表达，在其他组织中低表达。经相关分析表明：眼肉中 ucp3 基因的表达量与肌内脂肪和背膘厚关系不大，而与眼肌面积呈正相关（r=0.788）。结论：中国西门塔尔牛 ucp1、ucp2、ucp3 基因的表达，具有组织特异性；ucp1、ucp2、ucp3 在不同组织的表达差异，可能与其在不同组织中的功能有关；ucp3 基因可作为与胴体性状相关的候选基因。
孙国权
,
高树新
,
吴慧光
,
李俊雅
,
丽春
,
王景山
,
王玉泉
,
王国富
. 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
.
DOI: 10.7668/hbnxb.2014.04.019
The objective is to obtain expression profile of Chinese Simmental cattle ucp1,ucp2,ucp3 genes,to reveals the relationship between the ucp1,ucp2,ucp3 gene and energy metabolism,to explore the expression rule of key gene of fat deposition,and to screen the candidate gene affecting fat deposition.Total RNA were extracted from fifteen tissues(liver,lymph,intercostals muscle,shoulder muscle,spleen,kidney,rib eye muscle,lung,heart,gut fat,greater omental fat,testicle,small intestine,pancreas and backfat)of twenty animals from Chinese Simmental cattle.The primers of ucp1,ucp2,ucp3 genes were designed and synthesized,and the data were normalized using the β-actin gene as internal control.The specific expression of ucp1,ucp2,ucp3 genes in different tissues were investigated by real-time quantitative PCR.There were different degree expression of ucp1,ucp2,ucp3 genes mRNA in the tissues and organs of testicle,small intestine,heart,pancreas and so on.The expression of ucp1 was highest in the backfat,middle in the pancreas,small intestine,testicle,greater omental fat,gut fat,and heart,and lowest in the other tissues.The highest level expression of ucp 2 was detected in Testicle,and there were also higher expression of ucp2 in the lung,liver,and backfat than in other tissues.The highest level expression of ucp3 was in rib eye muscle,Intercostal muscle,and shoulder muscle,middle in the lung,and the lowest in other tissues.Correlation analyses showed that the expression level of ucp3 gene in rib eye had no significant relation with intramuscular fat and backfat thickness,but had positive relation to eye lean area(r= 0.788).It is concluded that the expression level of ucp1,ucp2,ucp3 genes had tissue specific might be related to the various functions on different tissues of cattle.The ucp3 gene can be used as a candidate gene related to carcass traits.
[1] Aysan E,Sahin F,Telci D,et al.Mechanism of body weight reducing effect of oral boric Acid intake[J].International Journal of Endocrinology,2013,3:914651.
[2] Yonezawa T,Sanosaka M,Haga S,et al.Regulation of uncoupling protein 2 expression by long-chain fatty acids and hormones in bovine mammary epithelial cells[J].Biochemical and Biophysical Research Communication,2008,375(2):280-285.
[3] Band M R,Larson J H,Rebeiz M, et al.An ordered comparative map of the cattle and human genomes[J].Genome Research, 2000,10(9):1359-1368.
[4] Winter A,van Eckevel M,Bininda-Emonds O R,et al.Genomic organization of the DGAT2/MOGAT gene family in cattle(Bos taurus)and other mammals[J].Cytogenetic and Genome Research,2003,102(1-4):42-47.
[5] Boss O,Samec S,Paolini-Giacobino A.Uncoupling protein-3:a new member of the mitochordrial carrier family with tissue-specific expression.[J].FEBS Letters,1997,408(1):39-42.
[6] Stone R T,Rexroad C E,Smith T P.Bovine UCP2 and UCP3 map to BTA15 [J].Animal Genetics,1999,30(5):378-381.
[7] Liu J,Li J,Li W J,et al.The role of uncoupling proteins in diabetes mellitus[J].Journal of Diabetes Research,2013:585-897.
[8] Raimbault S,Dridi S,Denjean F,et al.An uncoupling protein homologue putatively involved in facultative muscle thermogenesis in birds [J].The Biochemical Journal,2001,353(Pt3):441-444.
[9] Damon M,Vincent A,Lombardi A,et al.First evidence of uncoupling protein-2(UCP-2)and-3(UCP-3)gene expression in piglet skeletal muscle and adipose tissue[J].Gene,2000,246(1/2):133-141.
[10] Lin Y Q,Xu Y O,Yue Y,et al.Differences in mitochondrial gene expression profiles,enzyme activities and myosin heavy chain types in yak versus bovine skeletal muscles[J].Genetics and Molecular Research:GMR,2012,11(3):2871-2877.
[11] Zhang R,Li X.Association between IGF-IR,m-calpain and UCP-3 gene polymorphisms and growth traits in Nanyang cattle [J].Molecular Biology Reports, 2011,38(3):2179-2184.
[12] 刘永峰.牛13个体尺性状相关基因的克隆表达、序列特征及遗传效应分析[D].杨凌:西北农林科技大学,2010.
[13] Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T))Method[J].Methods.,2001,25(4):402-408.
[14] 乔永.湖羊羔羊不同部位肌肉肌内脂肪沉积相关基因表达的发育性变化研究[D].南京:南京农业大学,2007.
[15] 王宏博,梁春年,郭宪,等.欧拉型藏羊羔羊肌肉 UCP3 基因表达的研究[J].黑龙江畜牧兽医,2014(2):8-12,16.
[16] 彭广南.广西三黄鸡PPARγ和 UCP3 基因表达水平与脂肪性状的相关研究[D].南宁:广西大学,2013.
