猪解耦联蛋白基因3（UCP3）的序列及多态性研究
|
摘要
猪肉品质改良一直是家畜育种工作者工作的主要目标之一，随着现代人民生活水平的不断提高，生产质优量多的猪肉显得尤为重要。
本研究以与猪脂肪代谢有关的候选基因—解耦联蛋白基因3(UCP3)为研究对象，利用139头大白×梅山资源家系F_2代及321头F_1代资源家系(81头大梅、55头梅山、30头大白、43头长白、46头梅大、35头长大、31头大长)，研究了UCP3基因3’编码区及3’调控区的多态性和遗传效应，结果如下：
1、UCP3基因3’编码区的克隆测序及SNP检测---PCR-SSCP分析
根据猪UCP3基因的cDNA序列设计引物，对四个品种猪：大白、梅山、杜洛克、清平猪肌肉样提取总RNA进行反转录和特异性扩增，得到含UCP3基因3’编码区的扩增产物，经克隆及测序，对不同品种猪该区段序列进行比较分析，发现3个突变位点(SNPs)，选取其中一个(ORF内第842碱基C→T突变)能引起错义突变(甲硫氨酸→苏氨酸)的位点为标记位点进行标记基因型--性状的关联分析。
(1)利用PCR-SSCP在139头大白×梅山资源家系F_2代及321头F_1代资源家系(81头大梅、55头梅山、30头大白、43头长白、46头梅大、35头长大、31头大长)中进行了UCP3基因3’编码区的该标记位点基因型的分子扫描，发现：UCP3基因表现出3种基因型(AA、AB、BB)，结果显示：A等位基因频率在大白×梅山资源家系F_2代、大白×梅山资源家系F_1代个体及大白、长白、大长、长大群体中分布频率较高，其频率范围为0.71-0.87，而B等位基因则分布较低，其频率范围为0.13-0.29；在梅山、梅大两个群体中，B等位基因分布频率相对较高，分别是0.56、0.55。
(2)139头大白×梅山资源家系F_2代个体UCP3基因多态性与遗传效应分析—UCP3基因不同基因型对胴体性状的影响：
UCP3基因多态性与猪胸腰椎间膘厚极显著相关(P＜0.01)，UCP3~(AA)基因型个体胸腰椎间膘厚比UCP3~(AB)基因型个体低14.2％；同时UCP3基因多态性与猪眼肌高度、眼肌面积显著相关(P＜0.05)，UCP3~(AB)基因型个体眼肌高度、眼肌面积分别比UCP3~(BB)基因型个体高7.8％和11.2％。
在胴体性状方面，UCP3基因主要以加性作用方式发挥作用，眼肌高度、估测瘦肉率加性效应值分别为-0.348(P＜0.05)，-9.218(P＜0.01)。
(3)139头大白×梅山资源家系F_2代个体UCP3基因多态性与遗传效应分析—UCP3基因不同基因型对肉质性状的影响：
UCP3基因多态性与失水率、系水率及背最长肌色值显著相关，UCP3~(AA)基因型
猪解祸联蛋白基因3(UCP3)的序列及多态性分析
个体失水率比UCP3“B基因型个体低4.00，0(P<0.05)，其系水率则比ueP3“B基因型
个体高8.3%(P<0.05)，uCP3n日基因型个体背最长肌色值比ucP3从基因型个体高
18.2%(P<0.05):此外，UCP3基因多态性与水分(LD)极显著相关，UCP3AB基因
型个体水分比UCP3AA基因型个体低0.65%(P<0.01)。此外，在脂肪含量(LD)方
面，UcP3AB基因型猪比uCP3AA基因型个休高8.1%，达建议显著水平(P二0.067)。
在肉质性状方面，UCP3基因主要以加性作用方式发挥作用，显性效应不显著。
失水率、系水率及背最长肌色值的加性效应值分别为2.759(P<0.05)，一3.759
(P<0 .05)，2.012(P<0.05)。
(4)81头大白、梅山资源家系FI代个体UCP3基因多态性与遗传效应分析
一UCP3基因不同基因型对朋体性状的影响:
研究表明:UCP3基因多态性与肩部背膘厚相关显著:UCP3从基因型个体肩部
背膘厚比UCP3”“基因型个体高11 .3%(P<0.05);除此外，未发现在该家系中UcP3
不同基因型与其它性状的相关。
2 UCP3基因3’调控区Aval多态性位点的遗传效应分析
利用猪DNA对UCP3基因3’调控区的一碱基变异区段进行PCR扩增，对其扩
增产物的Aval酶切位点进行PCR一PFLP分析，并在1 39头大白又梅山参考家系FZ
代个体中进行多态性扫描和遗传效应分析，结果表明:该多态位点的不同基因型与
猪胭体及肉质性状相关均不显著。并且该位点基因除在水分性状(LD)中的加性效
应值显著外:一0.388(P<0.05)，其它性状的加性、显性效应也都不显著。
本实验结果表明:UCP3基因3’编码区突变位点的多态性与猪胭体性状:胸腰
椎间膘厚、眼肌高度、眼肌面积及肉质性状:失水率、系水率、背最长肌色值和水
分(LD)相关显著，同时与脂肪含量(LD)的相关达建议显著水平;并且，该基
因片段以加性效应为主，有利于在育种过程中的遗传固定。因此，建议是否将此标
记基因在猪标记辅助选择一肉质品质及胭休品质改良中加以应用。
Improvement of porcine meat quality is always one of the main goals for livestock genetics and breeding.At present.It is more important for technicians to produce pork with the lean and good quality,while people tend to be interested in meat quality when the standard of living condition has been improved.
In this paper,we selected the Uncoupling Protein Gene-3(UCP3) as the candidate gene for carcass and meat quality,and meanwhile analyzed its gene effects and the pattern of effects.
1. doing of UCP3 gene 3'-cDNA translated region and detection method of its single polymorphism(SNP)-PCR-SSCP
Using the porcine cDNA sequence.primers of UCP3 gene were designed and were applied to amplify the gene from four different porcine total RNA of longissimus dorsi muscle.Four breeds' fragments of 574bp length were got and we compared the differences of their sequences.The compared results showed that there were three single nucleotide mutations in UCP3 gene 3'-cDNA translated region, of which the mutation of 842bp in ORF can result in the change of ammo acid sequence.PCR-SSCP scanning of 139 F2 and 321 Fl individuals of pig resource families showed that:
(1) UCP3 gene showed three genotypes(AA,AB and BB).Their disstribution indicated that allele A seemed predominant in the following populations: Large Whitex Meishan F2 and Fl, Large White, Landrace, Large White Landrace, Landrace x Large White, and the ranges of allele frequencies was 0.71-0.87;in contrast.allele B was 0.13-0.29.However,the frequencies of allele B were 0.56 and 0.55 respectively.
(2) Association analyzing of UCP3 polymorphism with carcass traits in 139 F2 Large WhitexMeishan individuals
Significant association between the UCP3 genotypes and thorax-waist fat thickness,height and area of longissimus dorsi muscle(LD) was found.Pigs with the AA genotype had approximately 14.2% less average thorax-waist fat thickness than those with the AB genotype(P<0.01)and the height and area of longissimus dorsi muscle(LD) of BB genotype individuals were 7.8% and 11.2% less than AB genotype ones(P<0.05).
UCP3 gene played a role mainly in the pattern of additive effect on carcass traits.The additive effect values of longissimus dorsi muscle height and estimated lean meat percentage were -0.348(P<0.05) and -9.281 respectively(P<0.01).
(3) Association analyzing of UCP3 polymorphism with meat quality traits in 139 F2 Large WhitexMeishan individuals
It showed that UCP3 genotypes had significant association with water lost rate, water holding capacity, meat color(LD)(P<0.05), and water(LD) (P<0.01).The AA genotype individuals had 4.0%, 18.2% respectively less than BB genotype ones in water
lost rate, meat color, but had 8.3% more water holding capacity than pigs with BB genotype. To the water(LD),the AA genotype pigs were 0.65 more than AB ones.
We also found that UCP3 gene' additive effect on meat quality predominated in the population,and the additive effect values of water lost rate,water holding capacity,meat color(LD) were 2.750(P<0.05), -3.795(P<0.05), -2.012 (P<0.05) respectively.
(4)Scanning results of UCP3 polymorphism on carcass traits in 81 Fl Large WhitexMeishan resource family
Except for the significant association with shoulder fat thickness(P<0.05),we found no more significance in the association with other carcass traits. It showed that the BB genotype pigs were 11.3% less should fat thickness than AA genotype ones.
2. UCP3 gene 3'-UntransIated Region' polymorphism
The 696bp fragement of UCP3 gene 3'-Untranslated Region was gained after amplifying. Using Ava I , PCR-RFLP scanning in 139 F2 resource family indicated that there was no significant association of 3'-Untranslated Region' polymorphism with carcass traits and meat quality traits as well. However, the region of UCP3 gene seemed to play a mainly additive effect on water(LD):-0.388(P<0.05).
本研究以与猪脂肪代谢有关的候选基因—解耦联蛋白基因3(UCP3)为研究对象，利用139头大白×梅山资源家系F_2代及321头F_1代资源家系(81头大梅、55头梅山、30头大白、43头长白、46头梅大、35头长大、31头大长)，研究了UCP3基因3’编码区及3’调控区的多态性和遗传效应，结果如下：
1、UCP3基因3’编码区的克隆测序及SNP检测---PCR-SSCP分析
根据猪UCP3基因的cDNA序列设计引物，对四个品种猪：大白、梅山、杜洛克、清平猪肌肉样提取总RNA进行反转录和特异性扩增，得到含UCP3基因3’编码区的扩增产物，经克隆及测序，对不同品种猪该区段序列进行比较分析，发现3个突变位点(SNPs)，选取其中一个(ORF内第842碱基C→T突变)能引起错义突变(甲硫氨酸→苏氨酸)的位点为标记位点进行标记基因型--性状的关联分析。
(1)利用PCR-SSCP在139头大白×梅山资源家系F_2代及321头F_1代资源家系(81头大梅、55头梅山、30头大白、43头长白、46头梅大、35头长大、31头大长)中进行了UCP3基因3’编码区的该标记位点基因型的分子扫描，发现：UCP3基因表现出3种基因型(AA、AB、BB)，结果显示：A等位基因频率在大白×梅山资源家系F_2代、大白×梅山资源家系F_1代个体及大白、长白、大长、长大群体中分布频率较高，其频率范围为0.71-0.87，而B等位基因则分布较低，其频率范围为0.13-0.29；在梅山、梅大两个群体中，B等位基因分布频率相对较高，分别是0.56、0.55。
(2)139头大白×梅山资源家系F_2代个体UCP3基因多态性与遗传效应分析—UCP3基因不同基因型对胴体性状的影响：
UCP3基因多态性与猪胸腰椎间膘厚极显著相关(P＜0.01)，UCP3~(AA)基因型个体胸腰椎间膘厚比UCP3~(AB)基因型个体低14.2％；同时UCP3基因多态性与猪眼肌高度、眼肌面积显著相关(P＜0.05)，UCP3~(AB)基因型个体眼肌高度、眼肌面积分别比UCP3~(BB)基因型个体高7.8％和11.2％。
在胴体性状方面，UCP3基因主要以加性作用方式发挥作用，眼肌高度、估测瘦肉率加性效应值分别为-0.348(P＜0.05)，-9.218(P＜0.01)。
(3)139头大白×梅山资源家系F_2代个体UCP3基因多态性与遗传效应分析—UCP3基因不同基因型对肉质性状的影响：
UCP3基因多态性与失水率、系水率及背最长肌色值显著相关，UCP3~(AA)基因型
猪解祸联蛋白基因3(UCP3)的序列及多态性分析
个体失水率比UCP3“B基因型个体低4.00，0(P<0.05)，其系水率则比ueP3“B基因型
个体高8.3%(P<0.05)，uCP3n日基因型个体背最长肌色值比ucP3从基因型个体高
18.2%(P<0.05):此外，UCP3基因多态性与水分(LD)极显著相关，UCP3AB基因
型个体水分比UCP3AA基因型个体低0.65%(P<0.01)。此外，在脂肪含量(LD)方
面，UcP3AB基因型猪比uCP3AA基因型个休高8.1%，达建议显著水平(P二0.067)。
在肉质性状方面，UCP3基因主要以加性作用方式发挥作用，显性效应不显著。
失水率、系水率及背最长肌色值的加性效应值分别为2.759(P<0.05)，一3.759
(P<0 .05)，2.012(P<0.05)。
(4)81头大白、梅山资源家系FI代个体UCP3基因多态性与遗传效应分析
一UCP3基因不同基因型对朋体性状的影响:
研究表明:UCP3基因多态性与肩部背膘厚相关显著:UCP3从基因型个体肩部
背膘厚比UCP3”“基因型个体高11 .3%(P<0.05);除此外，未发现在该家系中UcP3
不同基因型与其它性状的相关。
2 UCP3基因3’调控区Aval多态性位点的遗传效应分析
利用猪DNA对UCP3基因3’调控区的一碱基变异区段进行PCR扩增，对其扩
增产物的Aval酶切位点进行PCR一PFLP分析，并在1 39头大白又梅山参考家系FZ
代个体中进行多态性扫描和遗传效应分析，结果表明:该多态位点的不同基因型与
猪胭体及肉质性状相关均不显著。并且该位点基因除在水分性状(LD)中的加性效
应值显著外:一0.388(P<0.05)，其它性状的加性、显性效应也都不显著。
本实验结果表明:UCP3基因3’编码区突变位点的多态性与猪胭体性状:胸腰
椎间膘厚、眼肌高度、眼肌面积及肉质性状:失水率、系水率、背最长肌色值和水
分(LD)相关显著，同时与脂肪含量(LD)的相关达建议显著水平;并且，该基
因片段以加性效应为主，有利于在育种过程中的遗传固定。因此，建议是否将此标
记基因在猪标记辅助选择一肉质品质及胭休品质改良中加以应用。
Improvement of porcine meat quality is always one of the main goals for livestock genetics and breeding.At present.It is more important for technicians to produce pork with the lean and good quality,while people tend to be interested in meat quality when the standard of living condition has been improved.
In this paper,we selected the Uncoupling Protein Gene-3(UCP3) as the candidate gene for carcass and meat quality,and meanwhile analyzed its gene effects and the pattern of effects.
1. doing of UCP3 gene 3'-cDNA translated region and detection method of its single polymorphism(SNP)-PCR-SSCP
Using the porcine cDNA sequence.primers of UCP3 gene were designed and were applied to amplify the gene from four different porcine total RNA of longissimus dorsi muscle.Four breeds' fragments of 574bp length were got and we compared the differences of their sequences.The compared results showed that there were three single nucleotide mutations in UCP3 gene 3'-cDNA translated region, of which the mutation of 842bp in ORF can result in the change of ammo acid sequence.PCR-SSCP scanning of 139 F2 and 321 Fl individuals of pig resource families showed that:
(1) UCP3 gene showed three genotypes(AA,AB and BB).Their disstribution indicated that allele A seemed predominant in the following populations: Large Whitex Meishan F2 and Fl, Large White, Landrace, Large White Landrace, Landrace x Large White, and the ranges of allele frequencies was 0.71-0.87;in contrast.allele B was 0.13-0.29.However,the frequencies of allele B were 0.56 and 0.55 respectively.
(2) Association analyzing of UCP3 polymorphism with carcass traits in 139 F2 Large WhitexMeishan individuals
Significant association between the UCP3 genotypes and thorax-waist fat thickness,height and area of longissimus dorsi muscle(LD) was found.Pigs with the AA genotype had approximately 14.2% less average thorax-waist fat thickness than those with the AB genotype(P<0.01)and the height and area of longissimus dorsi muscle(LD) of BB genotype individuals were 7.8% and 11.2% less than AB genotype ones(P<0.05).
UCP3 gene played a role mainly in the pattern of additive effect on carcass traits.The additive effect values of longissimus dorsi muscle height and estimated lean meat percentage were -0.348(P<0.05) and -9.281 respectively(P<0.01).
(3) Association analyzing of UCP3 polymorphism with meat quality traits in 139 F2 Large WhitexMeishan individuals
It showed that UCP3 genotypes had significant association with water lost rate, water holding capacity, meat color(LD)(P<0.05), and water(LD) (P<0.01).The AA genotype individuals had 4.0%, 18.2% respectively less than BB genotype ones in water
lost rate, meat color, but had 8.3% more water holding capacity than pigs with BB genotype. To the water(LD),the AA genotype pigs were 0.65 more than AB ones.
We also found that UCP3 gene' additive effect on meat quality predominated in the population,and the additive effect values of water lost rate,water holding capacity,meat color(LD) were 2.750(P<0.05), -3.795(P<0.05), -2.012 (P<0.05) respectively.
(4)Scanning results of UCP3 polymorphism on carcass traits in 81 Fl Large WhitexMeishan resource family
Except for the significant association with shoulder fat thickness(P<0.05),we found no more significance in the association with other carcass traits. It showed that the BB genotype pigs were 11.3% less should fat thickness than AA genotype ones.
2. UCP3 gene 3'-UntransIated Region' polymorphism
The 696bp fragement of UCP3 gene 3'-Untranslated Region was gained after amplifying. Using Ava I , PCR-RFLP scanning in 139 F2 resource family indicated that there was no significant association of 3'-Untranslated Region' polymorphism with carcass traits and meat quality traits as well. However, the region of UCP3 gene seemed to play a mainly additive effect on water(LD):-0.388(P<0.05).
引文
1．陈宏，蓝贤勇．分子标记及其应用．见：陈宏，耿社民主编，第八次全国畜禽遗传标记研讨会论文集，杨凌，2002：25-30
2．方宣钧，吴为人，唐纪良．作物DNA标记辅助育种．北京，科学出版社，2001
3．马月辉，吴常信，张沅．标记辅助选掸及其应用．曹红鹤，陈幼春主编，第六次全国畜禽遗传标记研讨会论文集，常州，1998．北京，中国人民出版社，1998，6：16-24
4 陈竺，强伯勤，方福德．基因组科学与人类疾病．北京，科学出版社，2001
5．蒋思文，彭健，熊远著．黑皮质索受体对动物采食量和能量稳态的调控．遗传，2002，24(2)：223-226
6．李凤娥．猪ESR和FSH-β位点对猪繁殖性状的调控机理研究．[博士学位论文]．武汉，华中农业大学，2002
7．鲁绍雄，吴常信．动物育种方法的回顾与展望．国外畜牧科技，2000，27(1)：24-28
8．屈彦纯。基因组扫描法定位猪1、2、6号染色体重要经济性状QTL的研究．[硕士学位论文]．武汉，华中农业大学，2002
9．熊远著，邓昌彦．种猪测定原理及方法．北京，中国农业出版社，1999b，57-118
10．赵兴波，方美英，李宁，吴常信．猪解耦联蛋白基因的克隆及多态性分析．中国动物遗传育种研究进展，2001：35-37
11．方美英，赵兴波，李宁．不同品种猪解耦联蛋白基因3’调控区的遗传变异．科学通报，2002，47(13),1010-1012．
12．罗怀容，施鹏，张亚平。单核苷酸多态性的研究技术。遗传，2001，23(5)：471-476
13．杨明。解耦联蛋白及其与肥胖的关系研究进展。国外医学内分泌学分册，2002，22(3)：169-172
14．郑文建。解耦联蛋白及其在基础代谢中的作用。国外医学．生理、病理科学与临床分册，2002，22(4)：418-420
15．张思仲。人类基因组的单核苷酸多态性及其医学应用。中华医学遗传杂志，1999，16：119-222
16. Andersson-Eklund L, Marklund L, Lundstrom K, Hsley C S, Andersson K, Hansson I,Moller M and Andersson L. Mapping Quantitative Trait Loci for Caeass and Meat Qulity Traits in a Wild Boar X Large White Intercross. J. Anim. Sci, 1998, 76:694-700
17. A Knoll, L Putnova, J Dvorak, G A Rohrer, S Cepica.Linkage mapping of an Ava Ⅰ PCR-RFLP within the porcine uncoupling protein 3 (UCP3) gene. Animal Genetics, 1999, 31:156-157
18. A. Lanni, L Bendeuee, A Lombardi, M Moreno, O Boss, P Muzzin, J P Giacobino, F Goglia. Expression of uncoupling proteins-3 and mitoehondrial activity in the
transition from hypothyroid to hyperthyroid state in rat skeletal muscle. FEBS, 1999, 444:250-254
19. Antonio Vidal-Puig, Gemma Solanes, Danica Grujic, Jeffrey S Flier, and Bradford B Lowell.UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue.Biochemical and Biophysical Research Communications, 1997, 235:79-82
20. Buetow K H,Edmonson M N,Cassidy A B.Reliable identification of large number of candidat SNPs from public EST data.Nat Genet, 1999, 21:323-325
21. Bostein D,White R L,Skolnick M,Davus R W.Constrution of a genetic linkage map in man using restricted fragment length polymorphisms.American Journal of Human Genetics, 1980,32:314-331
22. Bao S, Kennedy A, Wojciechowaki B, Liu YM.Expression of mRNAs encoding uncoupling proteins in human skeletal muscle: effect of obesity and diabetes[J]. Diabetes, 1998,47:1935-1940.
23. Campfield L A, Smith F J, Guisez Y, Devos R, Burn P. Recombinant mouse OB protein: evidence for a peripheral signal linking adiposity and central neutral networks. Science, 1995,269:546-549
24. Cheverud J M, Routman E J. Eiostasis and contribution to genetic variance components. Genetics, 1995,139:146-155
25. Da-wei Gong, Yufang He, and Marc, L.reitman. Genomic Organization and regulation by dietary fat of the uncoupling protein 3 and 2 gene.Biochemical and Biophysical Research Communications, 1999, 256:27-32
26. Da-Wei Gong, Yufang He, Marc L.Reitman, Michael Karas.uncoupling proteins-3 is a mediator of thermogenesis regulated by thyroid hormone, β3-adrenergic agonists,and leptin. The Journal of Biological Chemistry, 1997,39:24129-24132
27. Da-Wei Gong, Shadi Monemdjous, Oksana Gavrilova, Lisa R Leon.Lack of obesity and normal response to fasting and thyroid hormone in mice lacking uncoupling proteins-3. The Journalof Biological Chemistry, 2000,21: 16251-16257
28. Daniel Sanchis, Christophe Fleury, Nathalie Chomiki, Marc Goubern, Quinling Huang.BMCP1,a novel mitochondrial carrier with high expression in the central nervous system of humans and rodents,and respiration uncoupling activityin recombinant yeast.The Journal of Biological Chemistry, 1998,51:34611-34615
29. D E Moody, D Pomp,M K Nielsen and L D Van Vlak. Identification of quantitative trait loic related to energy balance in selection and inbred lines of mice. Genetics, 1999,152:699-711
30. Darvasi A, Soller M.Selective genotyping for determination of linkage between a marker and a quantitative traits locus. Genetics, 1992, 138:1365-1373
31. Eli Grindflek, Joanna Szyda, Zengting Liu and Sigbjorn Lien. Detection of quantitative trait loic for meat quality in a commercial slaught pig cross.
Mammalian Genome, 2001,12:299-304
32. Enerback S, Jadobsson A, Simpson EM, Guerra C.Mice lacking mitochondrial uncoupling protein are colding-sensitive but not obese. Nature, 1997,287:90-94.
33. Eshed Y, Zarmir D. An introgression line population of lycopersicon pennellii in the cultivated tomato enables the identification and fine mapping of yield-associted QTL.Genetics,1995, 141:1142-1162
34. Fabris R, Nisoli E, Lombardi AM, Chan CB.Preferential channeling of energy fuels towards fat rather than muscle during high free faty acid availability in rats.Diabetes, 2001,50(3):601-608.
35. Gemma Solanes, Antonio Vidal-puig, Danica Grujic, j Jeffrey S Flier.the human uncoupling proteins-3 gene.The Journal of Biological Chemistry, 1997, 41: 25433-25436
36. Hideki Yoshitomi,Kazuto Yarnazaki,Isao Tanaka.Cloing of mouse uncoupling proteins 3 cDNA and 5'-flanking region, and its genetic map. Gene, 1998,215:77-84
37. Hitoshi yamashita, yuzo sato, nozomu mori.different in induction of uncoupling protein gene in adipose tissues between young and old rats during cold exposure. FEBS Letters, 1999,458:157-161
38. Isabelle Cusin, Katerina E Zakrzewska, Olivier Boss, Patrick Muzzin. Chronic central leptin infusion enchances insulin-stimulated glucose metabolism and favors the expression of uncoupling proteins. Diabete, 1998,47:1014-1020
39. J.P.Bidanel and M.Rothschild. Current status of quantitative trait locus mapping in pigs. PigNews and Information, 2002, 23(2):39-5421
40. Kruglyak L.he uae of map of biallelic marks in linkage studies.Nature Gnetics, 1997,17:21-24
41. Karim S Echtary, Edith Winkler, Karina Frischmuth,and Martin Klingenberg. Uncoupling proteins 2 and 3 are highly active H~+ tranporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone). PNAS, 2001,4:1416-1421
42. Karim Salim Echtay, Cingyun Liucaskey, Edith Winkler, Karina Frischmuth. Regulation of UCP3 by nucleotides is different from regulation of UCPI. FEBS Letters, 1999,450:8-12
43. Karim S, Echtay, Edith Winkler & Martin Klingenberg.Conezyme Q is an obligatory cofactor for uncoupling protein function.Nature, 2000, 30:609-61
44. Knoll A, Putnova L, Dvorak J et al.Linkage mapping of an Ava Ⅰ PCR-RFLP within the porcine uncoupling protein 3(UCP3) gene. Anim. Genet., 2000,31(2):156-157.
45. Krook A, J Digby, A Orahilly, JR Zierath & H.Wallberg-Henriksson. Uncoupling protein 3 is reduced in skeletal muscle of NIDDM patients. Diabetes, 1998, 47:1528-1531.
46. Massoud Malek, Jack C M Dekkers, Hayyko K Lee, Tom J bass, Ken Prusa, Elisabeth Huff-Lonergan and Max F Rothchild.A molecular genome scan analysis to identify
chromosomal regions influencing economic traits in the pig. Ⅱ. Meat and muscle composition. Mammalian Genome, 2001,12:637-645
47. Mclaren D G, and Bovey M.Genetic influence on reproductive performance.VetClin N Am Food Anim Prac, 1992,8:435-439
48. Martin Jaburek, Miroslav Varecha,Ruth E Gimeno, Marlene Dembski, Petr Jezek. Transport function and regulation of mitochondrial uncoupling proteins 2 and 3. The Journal of Biological chemistry, 1999, 37:26003-26007
49. Mrie damon, annie vincent, assunta lombardi, patrick herpin.first evidence of uncoupling protein-2 (UCP2) and -3 (UCP3) gene expression in piglet skeletal muscle and adipose tissue. Gnene, 2000, 246:133-141
50. M Carmen Carmona, Angel Valmaseda,Sonia Brun, Octavi Vinas.Different regulation of uncoupling protein-2 and uncoupling protein-3 gene expression in brown adipose tissue during development and cold exposure.Biochemical and Biophysical Research Communications, 1998, 243:224-228
51. Millet L, Vidal H, Andreeli F.Increseaed uncoupling protein-2 and uncoupling protein-3 mRNA expression during fasting in obese and lean humans[J]. J Clinx Invest, 1997,100:2665-2670.
52. Nobuvc Tsubovama Kassoka,and Osamu Ezaki.Mitochondrial uncoupling protein 3 (UCP3) in skeletal muscle.Frotiers in Bioscience, 2001,6:570-574
53. Olivier Boss,Sonia Samec, Ariane Paoloni-Giacobino,Colette Rossier.Uncoupling protein-3:a new member of the mitochondrial carrier family with tissue-specific expression. FEB Letters, 1997,408:39-42
54. Oliver Bboss, Jean-Paul Giacobico, and Patrick Muzzin.Genomic structure of uncoupling protein-3 (UCP3) and its assignment to chromosome 11q13. Genomics, 1998,47:425-426
55. Oliver Bboss,soina samee,francoise kuhne,philippe bijlenga, uncoupling protein-3 expression in rodent skeletal muscle is modulated by food intake but not by changes in environmental temperature.the journal of biological chemistry. 1999,1:5-8
56. Petr Jezek, David E Orosz, Martin Mondriansky, and Keith D. Transport of anions and protons by the mitochondrial uncoupling protein and its regulation by nucleotides and fatty acids.The Journal of Biological Chemistry, 1994,42:26184-26190
57. Paszk A A, Dyas P A, Finn S T, Chen M, Buttram S T, Didion B A.Allelic effect of mico-satellites from putative QTL for reproductive traits.Anim Genet, 1998b 29(suppl1):69
58. P.Werner, S Neuenschwander, G Stranzinger.characterization of the porcine uncoupling proteins 2 and 3 (UCP2 & UCP3) and their localization to chromosome 9 p by somatic cell hybrids. Animal Genetics, 1999, 30:221-224
59. Rothschild M F, Jacobson C, Vaske D A, Tuggle C K, Short T H, Sasaki S, Eckardt G R and Mclaren D G,A major gene for litter size in pigs,Proc 5th World Congr
Genet.Appl Livest Prod,Guelph,Canada, 1994,21:225-228
60. Ruth E.Gimeno, Marlene Dembski, Xun Weng,Nanhua Deng.Cloing and characterization of an uncoupling protein homolog. Diabetes, 1997, 46:900-906
61 Ross PL, Lee L, Belgrader P. Discrinination of single-nucleotide polimorphisms in human DNA using peptide nucleicacid probes detected by MALDI-TOF mass spectrometry.Anal. Chemistry, 1997, 69:4197-4202
62. Short T H, Eckardt GR, Ssaski S, Rose M, Vincent A, Mclaren D G, Mileham A J, and Plastow G S.Marker assisted selection for litter size in pigs.J Anim Sci,1995,73 (suppl1): 109
63. Spinardi L, Mazars R, Theillet C.Protocol for an improved detection of point mutation by S SCP. Nucleic Acids Res, 1991, 19(14):4009
64. Splman R J and Bovenhuis H.Moving from QTL experimental results to the utilization of QTL in breeding program modles. Animal Genetics, 1998, 29:77-84
65. Sonia Brun, M Carmen Carmona, Teresa Mampel, Octavi Vinas. Activators of perixisome proliferator-activated receptor-α induce the expression of the uncoupling protein-3 gene in skeletal muscle.Diabete, 1999, 48:1217-1223
66. Thilo Hagen, Chen-yu Zhang, Lawrence J Slieker, Wendy K Chung. Assessment of uncoupling activity of the human uncoupling protein 3 short form and three mutants of uncoupling protein gene using yeast heterologous expression system. FEBS Letter, 1999454:201-207
67. Woods S C, Seely R J, Porte D Jr, Schwartz M W. Signal that regulate food intake and energy homeostasis.Science, 1998,280 (5368): 1378-1383
68 W Hinz, B Failer, S Gruninger, P Gazzotti,M Chiesi.ecombinant human uncoupling proteins-3 increase thermogensis in yeast cells. FEB S, 1999, 448: 57-61
69. Woods S C, Seely R J.Adiposity signals and the control of energy homeostasis [J].Nutrition, 20.00b,6(10):894-902
70. Y M Kennes, B D Murphy, F Pothier and M F Palin.Characterization of swine leptin(LEP) polymorphisms and their association with production traits.Animal Genetics, 32,215-218
71. Zhang YY, Riardo P, Margherita M. Positional cloing of the mouse obese gene and its human homologue. Nature, 1994,372:425-432.
2．方宣钧，吴为人，唐纪良．作物DNA标记辅助育种．北京，科学出版社，2001
3．马月辉，吴常信，张沅．标记辅助选掸及其应用．曹红鹤，陈幼春主编，第六次全国畜禽遗传标记研讨会论文集，常州，1998．北京，中国人民出版社，1998，6：16-24
4 陈竺，强伯勤，方福德．基因组科学与人类疾病．北京，科学出版社，2001
5．蒋思文，彭健，熊远著．黑皮质索受体对动物采食量和能量稳态的调控．遗传，2002，24(2)：223-226
6．李凤娥．猪ESR和FSH-β位点对猪繁殖性状的调控机理研究．[博士学位论文]．武汉，华中农业大学，2002
7．鲁绍雄，吴常信．动物育种方法的回顾与展望．国外畜牧科技，2000，27(1)：24-28
8．屈彦纯。基因组扫描法定位猪1、2、6号染色体重要经济性状QTL的研究．[硕士学位论文]．武汉，华中农业大学，2002
9．熊远著，邓昌彦．种猪测定原理及方法．北京，中国农业出版社，1999b，57-118
10．赵兴波，方美英，李宁，吴常信．猪解耦联蛋白基因的克隆及多态性分析．中国动物遗传育种研究进展，2001：35-37
11．方美英，赵兴波，李宁．不同品种猪解耦联蛋白基因3’调控区的遗传变异．科学通报，2002，47(13),1010-1012．
12．罗怀容，施鹏，张亚平。单核苷酸多态性的研究技术。遗传，2001，23(5)：471-476
13．杨明。解耦联蛋白及其与肥胖的关系研究进展。国外医学内分泌学分册，2002，22(3)：169-172
14．郑文建。解耦联蛋白及其在基础代谢中的作用。国外医学．生理、病理科学与临床分册，2002，22(4)：418-420
15．张思仲。人类基因组的单核苷酸多态性及其医学应用。中华医学遗传杂志，1999，16：119-222
16. Andersson-Eklund L, Marklund L, Lundstrom K, Hsley C S, Andersson K, Hansson I,Moller M and Andersson L. Mapping Quantitative Trait Loci for Caeass and Meat Qulity Traits in a Wild Boar X Large White Intercross. J. Anim. Sci, 1998, 76:694-700
17. A Knoll, L Putnova, J Dvorak, G A Rohrer, S Cepica.Linkage mapping of an Ava Ⅰ PCR-RFLP within the porcine uncoupling protein 3 (UCP3) gene. Animal Genetics, 1999, 31:156-157
18. A. Lanni, L Bendeuee, A Lombardi, M Moreno, O Boss, P Muzzin, J P Giacobino, F Goglia. Expression of uncoupling proteins-3 and mitoehondrial activity in the
transition from hypothyroid to hyperthyroid state in rat skeletal muscle. FEBS, 1999, 444:250-254
19. Antonio Vidal-Puig, Gemma Solanes, Danica Grujic, Jeffrey S Flier, and Bradford B Lowell.UCP3: an uncoupling protein homologue expressed preferentially and abundantly in skeletal muscle and brown adipose tissue.Biochemical and Biophysical Research Communications, 1997, 235:79-82
20. Buetow K H,Edmonson M N,Cassidy A B.Reliable identification of large number of candidat SNPs from public EST data.Nat Genet, 1999, 21:323-325
21. Bostein D,White R L,Skolnick M,Davus R W.Constrution of a genetic linkage map in man using restricted fragment length polymorphisms.American Journal of Human Genetics, 1980,32:314-331
22. Bao S, Kennedy A, Wojciechowaki B, Liu YM.Expression of mRNAs encoding uncoupling proteins in human skeletal muscle: effect of obesity and diabetes[J]. Diabetes, 1998,47:1935-1940.
23. Campfield L A, Smith F J, Guisez Y, Devos R, Burn P. Recombinant mouse OB protein: evidence for a peripheral signal linking adiposity and central neutral networks. Science, 1995,269:546-549
24. Cheverud J M, Routman E J. Eiostasis and contribution to genetic variance components. Genetics, 1995,139:146-155
25. Da-wei Gong, Yufang He, and Marc, L.reitman. Genomic Organization and regulation by dietary fat of the uncoupling protein 3 and 2 gene.Biochemical and Biophysical Research Communications, 1999, 256:27-32
26. Da-Wei Gong, Yufang He, Marc L.Reitman, Michael Karas.uncoupling proteins-3 is a mediator of thermogenesis regulated by thyroid hormone, β3-adrenergic agonists,and leptin. The Journal of Biological Chemistry, 1997,39:24129-24132
27. Da-Wei Gong, Shadi Monemdjous, Oksana Gavrilova, Lisa R Leon.Lack of obesity and normal response to fasting and thyroid hormone in mice lacking uncoupling proteins-3. The Journalof Biological Chemistry, 2000,21: 16251-16257
28. Daniel Sanchis, Christophe Fleury, Nathalie Chomiki, Marc Goubern, Quinling Huang.BMCP1,a novel mitochondrial carrier with high expression in the central nervous system of humans and rodents,and respiration uncoupling activityin recombinant yeast.The Journal of Biological Chemistry, 1998,51:34611-34615
29. D E Moody, D Pomp,M K Nielsen and L D Van Vlak. Identification of quantitative trait loic related to energy balance in selection and inbred lines of mice. Genetics, 1999,152:699-711
30. Darvasi A, Soller M.Selective genotyping for determination of linkage between a marker and a quantitative traits locus. Genetics, 1992, 138:1365-1373
31. Eli Grindflek, Joanna Szyda, Zengting Liu and Sigbjorn Lien. Detection of quantitative trait loic for meat quality in a commercial slaught pig cross.
Mammalian Genome, 2001,12:299-304
32. Enerback S, Jadobsson A, Simpson EM, Guerra C.Mice lacking mitochondrial uncoupling protein are colding-sensitive but not obese. Nature, 1997,287:90-94.
33. Eshed Y, Zarmir D. An introgression line population of lycopersicon pennellii in the cultivated tomato enables the identification and fine mapping of yield-associted QTL.Genetics,1995, 141:1142-1162
34. Fabris R, Nisoli E, Lombardi AM, Chan CB.Preferential channeling of energy fuels towards fat rather than muscle during high free faty acid availability in rats.Diabetes, 2001,50(3):601-608.
35. Gemma Solanes, Antonio Vidal-puig, Danica Grujic, j Jeffrey S Flier.the human uncoupling proteins-3 gene.The Journal of Biological Chemistry, 1997, 41: 25433-25436
36. Hideki Yoshitomi,Kazuto Yarnazaki,Isao Tanaka.Cloing of mouse uncoupling proteins 3 cDNA and 5'-flanking region, and its genetic map. Gene, 1998,215:77-84
37. Hitoshi yamashita, yuzo sato, nozomu mori.different in induction of uncoupling protein gene in adipose tissues between young and old rats during cold exposure. FEBS Letters, 1999,458:157-161
38. Isabelle Cusin, Katerina E Zakrzewska, Olivier Boss, Patrick Muzzin. Chronic central leptin infusion enchances insulin-stimulated glucose metabolism and favors the expression of uncoupling proteins. Diabete, 1998,47:1014-1020
39. J.P.Bidanel and M.Rothschild. Current status of quantitative trait locus mapping in pigs. PigNews and Information, 2002, 23(2):39-5421
40. Kruglyak L.he uae of map of biallelic marks in linkage studies.Nature Gnetics, 1997,17:21-24
41. Karim S Echtary, Edith Winkler, Karina Frischmuth,and Martin Klingenberg. Uncoupling proteins 2 and 3 are highly active H~+ tranporters and highly nucleotide sensitive when activated by coenzyme Q (ubiquinone). PNAS, 2001,4:1416-1421
42. Karim Salim Echtay, Cingyun Liucaskey, Edith Winkler, Karina Frischmuth. Regulation of UCP3 by nucleotides is different from regulation of UCPI. FEBS Letters, 1999,450:8-12
43. Karim S, Echtay, Edith Winkler & Martin Klingenberg.Conezyme Q is an obligatory cofactor for uncoupling protein function.Nature, 2000, 30:609-61
44. Knoll A, Putnova L, Dvorak J et al.Linkage mapping of an Ava Ⅰ PCR-RFLP within the porcine uncoupling protein 3(UCP3) gene. Anim. Genet., 2000,31(2):156-157.
45. Krook A, J Digby, A Orahilly, JR Zierath & H.Wallberg-Henriksson. Uncoupling protein 3 is reduced in skeletal muscle of NIDDM patients. Diabetes, 1998, 47:1528-1531.
46. Massoud Malek, Jack C M Dekkers, Hayyko K Lee, Tom J bass, Ken Prusa, Elisabeth Huff-Lonergan and Max F Rothchild.A molecular genome scan analysis to identify
chromosomal regions influencing economic traits in the pig. Ⅱ. Meat and muscle composition. Mammalian Genome, 2001,12:637-645
47. Mclaren D G, and Bovey M.Genetic influence on reproductive performance.VetClin N Am Food Anim Prac, 1992,8:435-439
48. Martin Jaburek, Miroslav Varecha,Ruth E Gimeno, Marlene Dembski, Petr Jezek. Transport function and regulation of mitochondrial uncoupling proteins 2 and 3. The Journal of Biological chemistry, 1999, 37:26003-26007
49. Mrie damon, annie vincent, assunta lombardi, patrick herpin.first evidence of uncoupling protein-2 (UCP2) and -3 (UCP3) gene expression in piglet skeletal muscle and adipose tissue. Gnene, 2000, 246:133-141
50. M Carmen Carmona, Angel Valmaseda,Sonia Brun, Octavi Vinas.Different regulation of uncoupling protein-2 and uncoupling protein-3 gene expression in brown adipose tissue during development and cold exposure.Biochemical and Biophysical Research Communications, 1998, 243:224-228
51. Millet L, Vidal H, Andreeli F.Increseaed uncoupling protein-2 and uncoupling protein-3 mRNA expression during fasting in obese and lean humans[J]. J Clinx Invest, 1997,100:2665-2670.
52. Nobuvc Tsubovama Kassoka,and Osamu Ezaki.Mitochondrial uncoupling protein 3 (UCP3) in skeletal muscle.Frotiers in Bioscience, 2001,6:570-574
53. Olivier Boss,Sonia Samec, Ariane Paoloni-Giacobino,Colette Rossier.Uncoupling protein-3:a new member of the mitochondrial carrier family with tissue-specific expression. FEB Letters, 1997,408:39-42
54. Oliver Bboss, Jean-Paul Giacobico, and Patrick Muzzin.Genomic structure of uncoupling protein-3 (UCP3) and its assignment to chromosome 11q13. Genomics, 1998,47:425-426
55. Oliver Bboss,soina samee,francoise kuhne,philippe bijlenga, uncoupling protein-3 expression in rodent skeletal muscle is modulated by food intake but not by changes in environmental temperature.the journal of biological chemistry. 1999,1:5-8
56. Petr Jezek, David E Orosz, Martin Mondriansky, and Keith D. Transport of anions and protons by the mitochondrial uncoupling protein and its regulation by nucleotides and fatty acids.The Journal of Biological Chemistry, 1994,42:26184-26190
57. Paszk A A, Dyas P A, Finn S T, Chen M, Buttram S T, Didion B A.Allelic effect of mico-satellites from putative QTL for reproductive traits.Anim Genet, 1998b 29(suppl1):69
58. P.Werner, S Neuenschwander, G Stranzinger.characterization of the porcine uncoupling proteins 2 and 3 (UCP2 & UCP3) and their localization to chromosome 9 p by somatic cell hybrids. Animal Genetics, 1999, 30:221-224
59. Rothschild M F, Jacobson C, Vaske D A, Tuggle C K, Short T H, Sasaki S, Eckardt G R and Mclaren D G,A major gene for litter size in pigs,Proc 5th World Congr
Genet.Appl Livest Prod,Guelph,Canada, 1994,21:225-228
60. Ruth E.Gimeno, Marlene Dembski, Xun Weng,Nanhua Deng.Cloing and characterization of an uncoupling protein homolog. Diabetes, 1997, 46:900-906
61 Ross PL, Lee L, Belgrader P. Discrinination of single-nucleotide polimorphisms in human DNA using peptide nucleicacid probes detected by MALDI-TOF mass spectrometry.Anal. Chemistry, 1997, 69:4197-4202
62. Short T H, Eckardt GR, Ssaski S, Rose M, Vincent A, Mclaren D G, Mileham A J, and Plastow G S.Marker assisted selection for litter size in pigs.J Anim Sci,1995,73 (suppl1): 109
63. Spinardi L, Mazars R, Theillet C.Protocol for an improved detection of point mutation by S SCP. Nucleic Acids Res, 1991, 19(14):4009
64. Splman R J and Bovenhuis H.Moving from QTL experimental results to the utilization of QTL in breeding program modles. Animal Genetics, 1998, 29:77-84
65. Sonia Brun, M Carmen Carmona, Teresa Mampel, Octavi Vinas. Activators of perixisome proliferator-activated receptor-α induce the expression of the uncoupling protein-3 gene in skeletal muscle.Diabete, 1999, 48:1217-1223
66. Thilo Hagen, Chen-yu Zhang, Lawrence J Slieker, Wendy K Chung. Assessment of uncoupling activity of the human uncoupling protein 3 short form and three mutants of uncoupling protein gene using yeast heterologous expression system. FEBS Letter, 1999454:201-207
67. Woods S C, Seely R J, Porte D Jr, Schwartz M W. Signal that regulate food intake and energy homeostasis.Science, 1998,280 (5368): 1378-1383
68 W Hinz, B Failer, S Gruninger, P Gazzotti,M Chiesi.ecombinant human uncoupling proteins-3 increase thermogensis in yeast cells. FEB S, 1999, 448: 57-61
69. Woods S C, Seely R J.Adiposity signals and the control of energy homeostasis [J].Nutrition, 20.00b,6(10):894-902
70. Y M Kennes, B D Murphy, F Pothier and M F Palin.Characterization of swine leptin(LEP) polymorphisms and their association with production traits.Animal Genetics, 32,215-218
71. Zhang YY, Riardo P, Margherita M. Positional cloing of the mouse obese gene and its human homologue. Nature, 1994,372:425-432.