蒙古羊和滩羊H-FABP基因、PPAR-γ基因与IMF的变化规律及其相关性研究
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摘要
近年来,世界各国的肉品消费量越来越大,而且已经出现一种由单纯的重视数量向重视质量方向发展。所以,畜牧生产应该顺应这种趋势,对生产方式方法进行相应的调整,以追求更高的社会效益和经济效益。
本研究针对当前羊肉品质存在的品质不理想的问题,以肌内脂肪含量为突破口,从心脏型脂肪酸结合蛋白(H-FABP)基因和过氧化物酶体增殖物激活受体γ(PPAR-γ)基因mRNA的表达量出发,探讨其在我国两个绵羊品种中的表达量变化趋势以及其对羊肉肌内脂肪含量的影响。力求为今后的育种工作提供一定的理论基础。
选取相同饲养条件的滩羊和蒙古羊各12只分别于90、120、160和200日龄进行屠宰。用半定量RT-PCR方法分别测定每个个体背最长肌中的心脏型脂肪酸结合蛋白(H-FABP)基因和过氧化物酶体增殖物激活受体γ(PPARγ)基因的相对表达含量及其肌内脂肪含量(IMF),分析其发育规律及其之间的相关性。结果表明:
(1)滩羊IMF第二阶段比第一阶段含量高,第二阶段之后变化不明显,第一阶段和其它各阶段差异显著(P<0.05);
(2)蒙古羊随日龄增加,肌内脂肪含量不断升高,但是各阶段差异不显著;
(3)两个品种绵羊的H-FABP基因相对表达量随着日龄升高而升高,在160日龄达到最高后开始下降。其中,滩羊160日龄与前两阶段差异显著(P<0.05),蒙古羊160日龄与其它三阶段均差异显著(P<0.05);
(4)滩羊PPAR-γ基因相对表达含量变化规律不明显,但在160日龄相对表达量最高,与其它阶段差异显著(P<0.05)。蒙古羊随着日龄增加而降低,在160日龄后开始升高,各阶段差异不显著;
(5)蒙古羊第一阶段到第三阶段(90-160日龄)期间H-FABP基因与IMF相关系数为0.988(P=0.068),蒙古羊第一阶段到第三阶段(90-160日龄)期间PPAR-γ基因与IMF相关系数为-0.8849(P=0.1084)。
通过本试验我们得知,在一定生长时期内,我国某些地方品种中,一些重要的影响肉质性状的基因对其肌内脂肪的影响特别明显,应该引起育种工作者的高度重视。
In recent years, the consumption of meat in the world is improved swiftly and violently, and the quality is more regard than quantity. So the production of stockbreeding should improve for this demand, to adjust the need of Social and Economic benefits.
To improve the situation about low quality in sheep mutton, the study is designed for the direction of express level about mRNA of H-FABP and PPARγof intramuscular fat acid content. The Rule and Relativity on IMF Content of H-FABP and PPAR-γmRNA Expression Level are detected, and hope to provide some basic research for the future.
12 Tan Sheep and 12 Meng Gu Sheep breeds in same condition were killed on Day 90, Day 120, Day 160 and Day 200. The level of expression of H-FABP and PPAR-γin Longissimus dorsal muscle was detected by the method of Semi-Quantitative RT-PCR Assay, and the IMF content was detected to study the rule between the growth and the relativity of the IMF content at the expression level of gene. The results as follows:
(1)The IMF content in Tan Sheep of the second period was higher than the first, and no obvious change later then, the first period shows significant differences to the others(P<0.05);
(2)The IMF content of Meng Gu Sheep was ascend with the days, but there is no significant differences between 4 periods;
(3)The expression of H-FABP in two breeds was ascended with days until Day 160, and then it began to go down. The expression of Tan Sheep on Day 160 takes a significant difference to the first and the second periods. The expression of Meng Gu Sheep on Day 160 was significant differences to the other 3 periods;
(4)There is no obvious rule in the expression level of PPAR-γof Tan Sheep. It was highest on Day 160, which showed significant differences to the other periods. The Meng Gu Sheep was declined with the time until Day 160, and began to descend;
(5) In Meng Gu sheep, the mRNA expression level of H-FABP was highly positively correlated (r=0.988, P=0.068) with IMF content from Day 90 to 160, but that of PPAR-γwas highly negatively correlated (r=-0.8849, P=0.1084) with IMF content in the same period.
本研究针对当前羊肉品质存在的品质不理想的问题,以肌内脂肪含量为突破口,从心脏型脂肪酸结合蛋白(H-FABP)基因和过氧化物酶体增殖物激活受体γ(PPAR-γ)基因mRNA的表达量出发,探讨其在我国两个绵羊品种中的表达量变化趋势以及其对羊肉肌内脂肪含量的影响。力求为今后的育种工作提供一定的理论基础。
选取相同饲养条件的滩羊和蒙古羊各12只分别于90、120、160和200日龄进行屠宰。用半定量RT-PCR方法分别测定每个个体背最长肌中的心脏型脂肪酸结合蛋白(H-FABP)基因和过氧化物酶体增殖物激活受体γ(PPARγ)基因的相对表达含量及其肌内脂肪含量(IMF),分析其发育规律及其之间的相关性。结果表明:
(1)滩羊IMF第二阶段比第一阶段含量高,第二阶段之后变化不明显,第一阶段和其它各阶段差异显著(P<0.05);
(2)蒙古羊随日龄增加,肌内脂肪含量不断升高,但是各阶段差异不显著;
(3)两个品种绵羊的H-FABP基因相对表达量随着日龄升高而升高,在160日龄达到最高后开始下降。其中,滩羊160日龄与前两阶段差异显著(P<0.05),蒙古羊160日龄与其它三阶段均差异显著(P<0.05);
(4)滩羊PPAR-γ基因相对表达含量变化规律不明显,但在160日龄相对表达量最高,与其它阶段差异显著(P<0.05)。蒙古羊随着日龄增加而降低,在160日龄后开始升高,各阶段差异不显著;
(5)蒙古羊第一阶段到第三阶段(90-160日龄)期间H-FABP基因与IMF相关系数为0.988(P=0.068),蒙古羊第一阶段到第三阶段(90-160日龄)期间PPAR-γ基因与IMF相关系数为-0.8849(P=0.1084)。
通过本试验我们得知,在一定生长时期内,我国某些地方品种中,一些重要的影响肉质性状的基因对其肌内脂肪的影响特别明显,应该引起育种工作者的高度重视。
In recent years, the consumption of meat in the world is improved swiftly and violently, and the quality is more regard than quantity. So the production of stockbreeding should improve for this demand, to adjust the need of Social and Economic benefits.
To improve the situation about low quality in sheep mutton, the study is designed for the direction of express level about mRNA of H-FABP and PPARγof intramuscular fat acid content. The Rule and Relativity on IMF Content of H-FABP and PPAR-γmRNA Expression Level are detected, and hope to provide some basic research for the future.
12 Tan Sheep and 12 Meng Gu Sheep breeds in same condition were killed on Day 90, Day 120, Day 160 and Day 200. The level of expression of H-FABP and PPAR-γin Longissimus dorsal muscle was detected by the method of Semi-Quantitative RT-PCR Assay, and the IMF content was detected to study the rule between the growth and the relativity of the IMF content at the expression level of gene. The results as follows:
(1)The IMF content in Tan Sheep of the second period was higher than the first, and no obvious change later then, the first period shows significant differences to the others(P<0.05);
(2)The IMF content of Meng Gu Sheep was ascend with the days, but there is no significant differences between 4 periods;
(3)The expression of H-FABP in two breeds was ascended with days until Day 160, and then it began to go down. The expression of Tan Sheep on Day 160 takes a significant difference to the first and the second periods. The expression of Meng Gu Sheep on Day 160 was significant differences to the other 3 periods;
(4)There is no obvious rule in the expression level of PPAR-γof Tan Sheep. It was highest on Day 160, which showed significant differences to the other periods. The Meng Gu Sheep was declined with the time until Day 160, and began to descend;
(5) In Meng Gu sheep, the mRNA expression level of H-FABP was highly positively correlated (r=0.988, P=0.068) with IMF content from Day 90 to 160, but that of PPAR-γwas highly negatively correlated (r=-0.8849, P=0.1084) with IMF content in the same period.
引文
[1] Novakofski J.Adipogenesis.Usefulness of in vitro and in vivo experiment models.Journal of Animal Science,2004,82:905~915.
[2] 陈润生.优质猪肉的指标及其度量方法[J].东北农业大学学报.
[3] 吴 伟,孙飞舟,胡勇军.百日龄羔羊肉肉质分析测定[J].中国草食动物,2000,2(2):32~33.
[4] 陈代文,张克英,胡祖禹.猪肉品质特征的形成原理[J].四川农业大学学报,2002,20(1): 60~66.
[5] 胡宝利.不同年龄秦川牛胴体性状与肉质性状的研究[D].陕西:西北农业大学,2001
[6] Brennand,C.P.Ha J.K.and Lindsay,R.C.Aroma Properties and Thresholds of Some Branched-chain and otherminor volatile fatty acid soccurring in milk fat meat lipids[J].J.Sensory Stud.1989,4:105~120
[7] Vignal A,Milan D,San Cristobal M, et al.A review on SNP and other types of molecular markers and their use in animal genetics[J].Genetics Selection Evolution,2002’34:275~305.
[8] 王悦生等。金华猪肉品质的研究[J].浙江农业大学学报,1982.8.
[9] WilliamsJL.The impact of marker-assisted selection in animal breeding and biotechnology [J].Scientific and Technical Review,2005,24:379~391.
[10] Womack J E.The impact of sequencing the bovine genome[J].Australian Journal of Experimental Agriculture,2006,46(2):151~153.
[11] Anderson H J.What is pork quality[A].Inc:Quality of meat and fat in pigs as affected by genetic and nutrition.
[12] 张伟力.现代猪肉品质概念与检测手段进展[J].养猪,2004,5:45～47
[13] Roschnik R.New trends in consumer acceptant and convenient[A].In:Proceedings of EuroFood ChemV,Ver sailles[C].France,1989,30～39.
[14] Novakofski J.Adipogenesis: Usefulness of in vitro and in vivo experiment models.Journal of Animal Science,2004,82: 905~915.
[15] Hodgson R R,Davis G W,Smith G C,Savell J W,Cross H R.Relationships between pork loin palatability traits and physical characteristics of cooked chops.Journal of Animal Science,1991,69:4858~4865.
[16] Wang Y H,Byrne K A,Reverter A,Harper G S,Taniguchi M,McWilliam S M,Mannen H,Oyama K,Lehnert S A.Transcriptional profiling of skeletal muscle tissue from two breeds of cattle.Mammalian Genome,2005,16: 201~210.
[17] 李凤娜,王继成,陈晓安.猪肉品质及其影响因素的研究概况[J].畜禽业.2006,6(199):12~15.
[18] Monin G,Ouali A.Muscle dierentiation and meat quality [M].InR Lawrie (Ed.): Developments in meat science (5thed),1991,(pp.89~157).
[19] Karlsson A,Enfalt A C,Essen-Gustavsson B,et al.Muscle histochemical and biochemical properties in relation to meat quality during selection for increased lean tissue growth rate in pigs [J].J Anim Sci,1993,71: 930~938.
[20] OHalloran G R,Troy D J,Buckley D J,et al.The role of endogenous proteases in the tenderization of fast glycolysing muscle [J].Meat Sci,1997,47: 187~121.
[21] Renand G,Picard B,Touraille C,et al.Relationships between muscle characteristics and meat quality traits of young Charolais bulls [J].Meat Sci,2001,59: 49~60.
[22] 孙 好 学 , 田 云 , 何 宏 轩 等 . 肌 内 脂 肪 酸 影 响 因 素 和 侯 选 基 因 研 究 进 展 [J]. 动 物 医 学 进展.2006,27(11):49~53.
[23] Warriss P D.Meat Science:An Introductory Text[M].CABI Publishing,2000.
[24] LawrieR A,Meat Science[M].Pergamon Press,1991.
[25] 黄启超.猪肉脂肪概念及其测定方法研究[J].食品科技.2006,8:134~139.
[26] 徐宁迎,赵兴波,蒋思文.猪鸡肉质性状分子标记及主效基因的研究进展[J].中国畜牧杂志,2004,4: 42~44.
[27] Farmer L J.The role of nutrients in meat ?avour formation [J].Proceedings of the Nutrition Society,1994,53: 327~333
[28] Liu Xiao-yan.Studies on Polymorphisms of H–FABP Gene and the Association With Intra muscular Fat Content in Pigs[D]
[29] HUANG Zhi-Guo,XIONG Li,LIU Zhen-Shan,The Developmental Changes and Effect on IMF Content of H-FABP and PPARγ mRNA Expression in Sheep Muscle[J].Acta Genetica Sinica,June 2006,33 (6):507~514.
[30] Williams J L.The use of marker-assisted selection in animal breeding and biotechnology [J].Scientific and Technical Review,2005,24: 379~391.
[31] wustone456.什么是候选基因.http://zhidao.baidu.com/question/41361388.html.2007-12-10 17:01
[32] Vignal A,Milan D,San Cristobal M,et al.A review on SNP and other types of molecular markers and their use in animal genetics [J].Genetics Selection Evolution,2002,34: 275~305.
[33] Veerkamp J H,van Moerkerk H B T,Prinsen C F M,et al.Structural and functional study on different human FABP types [J].Molecular and Cellular Biochemistry,1999,192: 137~142.
[34] RNA提取和RT-PCR.http//www.biorule.com,service@biorule.com.2007-4-17 20:33:34.
[35] Ockner RK.Histone overview of studies on fatty acid-binding proteins[J].Mol Cell Biochem.1990,98(1-2):3~9.
[36] Reese-Wagoner A,Thompson J,Banaszak L.Structural properties of the adipocyte lipid binding protein [J].Biochimica et Biophysica Acta,1999,1441: 106~116.
[37] Gerbens F,Jansen A,van Erp AJ,et al.The adipocyte fatty acid-binding protein locus:characterization and association with intramuscular fat content in pigs[J].Mamm Genome.1998,9(12):1022~1026
[38] Likic V A,Juranic N,Macuea S,et al.A structural water molecule in the family of fatty acid binding proteins [J].Prot Sci,2000,9: 497~504
[39] óvilo C,Pérez-Enciso M,Barragán C,et al.A QTL for intramuscular fat and backfat thickness is located on porcine chromosome 6 [J].Mamm Genome,2000,11: 344–346
[40] 叶满红.鸡脂肪酸结合蛋白基因的克隆及其与肌内脂肪含量的关系[D].2003,6
[41] Fischer H,Gustafsson T,Sundberg C J,et al.Fatty acid binding protein 4 in human skeletal muscle [J].BBRC,2006,346: 125~130
[42] 张 桂 香 ,9 个 猪 种 H-FABP 基 因 5’- 上 游 区 和 第 二 内 含 子 的 遗 传 变 异 [J]. 畜 牧 兽 医 学报,2002,33(4):304~343
[43] 李 桢 ; 曹 红 鹤 ; 储 明 星 . 中 外 11 个 猪 种 H-FABP 基 因 PCR-RFLP 的 研 究 [J]. 畜 牧 兽 医 学报,2003,34(4),313~317.
[44] Nechtellberger D,Pires V,Soolknet J,et al.Intramuscular fat content and genetic variants at fatty acid-binding protein loci in Austrian pigs[J].Anim Sci,2001,79(11):2798~2804.
[45] 陈敏.DHEA、运动通过PPARα对大鼠肝脏、心脏及肌肉中脂肪分解的调节作用[D].
[46] Lemberger T ,B Desvergne ,W Wahli .Peroxisome proliferator-activated receptors : A nuclear receptor signaling pathway in lipid physiology[J].Annu.Rey.Cell Dev.Biol,1996 ,12 :335～363.
[47] Schoonjans K,B Staels ,J Auwerx ,Role of the peroxisome proliferator-activated receptor (PPAR) in mediating the effects of fibrates and fatty acids on gene expression[J].Lip.Res.,1996 ,37 :907～925.
[48] Isabelle Issemann ,Stephen Green.Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators[J].Nature ,1990,347:645～650.
[49] 孟和,李辉,王宇祥.鸡PPARs基因组织表达特性的研究[J].遗传学报.2004,31(7):682~687.
[50] H.Sundvold,A.Brzozowska,and S.Lien. Characterisation of Bovine Peroxisome Proliferator-Activated Receptorsyγ1 and γ2:Genetic Mapping and Diferential Expression of the Two isoforms [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,1997,239,857-861.
[51] Mangelsdorf DJ,Thrmmel C,Beato M,et a1.The nuclear receptor superfamily:the second decade.Cell 1995;83:835-83.
[52] 张乾勇.PPAR的结构与功能及其生物学作用[J].国外医学卫生学分册,2000;27(5):284~288.
[53] Elangbam C.S,Tyler R D,Lightfoot RM.Peroxisome proliferator-activated receptors in atherosclerosis and inflammation an update[J].Toxicol Patho1 2001;29(2):224~231.
[54] Braissant O,Foufelle F,Scoto C,et a1.Differential expression of peroxisome proliferator-activated receptors(PPARs):tissue distribution of PPAR-alpha,-beta,and -gamma in the adult rat.Endocrinology 1996;137:354-366.
[55] Mattew E ,Poynter, Daynes R ATeroxisome proliferator-activated receptor a activation modulates cellular redox status,represses nuclear factor-kB signaling,and reduces inflammatory cytokine production in aging.J Biol Chem 1998;273(49):32833-32841
[56] Moras D, Gronemeyer H. The nuclear receptor ligand binding domain:st ructure and function. CurrO pin Cell Biol,1998 ,10 :381391
[57] Devchand PR,Keller H,Peters JM,et al.The PPAR a leukotriene B4 pathway to inflammation control.Nature 1996;384:39-43
[58] Martin NF ,Furman C ,D ele riv e Pet a1.Peroxisome proliferator-activated receptor activators inhibit oxidized low-density lipoprotein-induced endothelin-1 secretion in endothelial cells.J Cardiovasc Pharmacol 2002;40(6):822-831
[59] Elangbam C S, Tyler R D, Light foot RM.Peroxisome proliferator-activated receptors in atherosclerosis and inflammation an update.ToxicolPa tho12001;29(2):224-231.
[60] Stacls.B ,Wolfgang K, Habib A, et a1.Activation of human aortic smooth-muscle cells is inhibited by PPAR a but not PPARγactivators. Nature 1998;393:790-793
[61] Fajias L,Auboeuf D.The organization,promoter analysis,and expression of the human PPARγgene[J].The Journal of Bio-logical Chemistry,1997,272(30):18779-18789.
[62] 孟和 , 王桂华 , 王启贵 . 鸡 PPAR 基因单核苷酸多态与脂肪性状相关的研 究 [J]. 遗传学报,2002,29(2):119~123.
[63] Tontonoz P E Hu,B M Spoegelman.Stimulation of adipogenesis in fibroblasts by PPARγ2,a lipid-activated transcription factor[J].Cell,1994,79:1147~1156.
[64] 罗敏,左祥生.核转录因子PPARγ与肥胖、糖尿病的临床联系及进展[J].辽宁实用糖尿病杂志2001,9 (2):49~51Dreyer C H Keller,A Mahfoudi,V Laudet,G Krey,W Wahli Positive,Regulation of theperoxisomal betaoxidation pathway by fatty acids through activation of peroxisomal proliferator-activated receptors(PPAR)[J].Biol.Cell,1993,77:67~76
[65] Hertzel A V,Bennaars-Eiden A,Bernlohr D A.Increased lipolysis in transgenic animals over expressing the epithelial fatty acid binding protein in adipose cell [J].Lipid Res,2002,43(12): 2105~2111.
[66] Lim H,Gupta R,Ma Wet al.Cycloxygenase-2 derived prostacyclin mediates embryo implantation in the mouse via PPARδ[J].Genes Dev1999;13:1561一1574.
[67] Foman BM, Tontonoz P, Chen J, et a1. 15-Deoxy-12,14 prostaglandin J2 is a ligand for the adipocyte determination Prostaglandin factor PPAR[J].Cell,19 95,83 :80 3-812.
[68] Issemann.L .and Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators[J].Nature,1990,34 7(6295):64 5-650
[69] HUANG Zhi-Guo,XIE Zhuang.The research development of livestock and poultry adipose related gene[J].Animal Husbandry & Veterinary Medicine,2004,36(4):41~43.
[70] C Diot ,MDuaire.Characterization of a cDNA sequence encoding the peroxisome proliferator- activated receptor α in the chicken[J].Poultry Science.1999,78:1198～1202.
[71] Evans R M.The steroid and thyroid hormone receptor superfamily [J ].Sc ience,19 88,24 0:88 9-895.
[72] Chawla A, Schwarz E J, et al. Peroxisome proliferator-activated receptor (PPAR) gamma:adi pos e-p re dom inantex pressionan din ductione arlyin a dipocyted iferentiation[J ].En docrinology,199 4, 13 5( 2): 79 8-800.
[73] 姜俊芳.脂肪细胞分化相关因子基因表达在猪生长过程中的变化规律研究[D] 杭州:浙江大学,2006
[74] Xu HE,Lambert MH,Valcric G,et al.Moleecular recognition of fatty acidsby perocisome proliferators-activated receptors[J].Mo Cell,199,3(3):397-403.
[75] RNA 提取策略及RT-PCR技术.http//www.biorule.com,service@biorule.com
[76] Bligh,E,G,.Dyer,W,J.A rapid method of total lipid extraction and purification and purification[j].Can.Biochem Physiol.1959(37):911~917
[77] May J,Costa M,Ojeda S R.Developmental expression of the genes encoding transforming growth factor alpha and its receptor in the by prothalamus of female thesus macaques[J].Neurcendocrino, 1994, 60(1):346.
[78] Stanley K K,Szewczuk E.Multiplexed tandem PCR: gene profiling from small amounts of RNA using SYBR Green detection [J].Nucleic Acids Research,2005,33(20): e180
[79] 徐春兰,汪以真.半定量RT-PCR法分析猪肌肉组织细胞谷胱甘肽过氧化物酶mRNA表达水平[J].中国兽药杂志.2005,39(8):3~6;23.
[80] 陈昕,王保莉,曲东,等.小麦硫转运蛋白基因半定量RT-PCR检测方法的建立[J].西北植物学报,2006,26(2):0309~0313.
[81] Chen D,Klebe R J.Controls for validation forrelative reverse transcription-polymerase chain reaction assays.PCR Methods Appl,1993,3:127~129.
[82] Marone M,scambia G,Giannitelli C,et al.Cyclin E and cdk2 alterations in ovarian cancer: Amplification and overexpression .Int J Cancer,1997,74:390~395
[83] FreemanW M,WalkerS J,VranaK E.Quantitative RT-PCR:Pitfalls and potential[ J].BioTechniaues, 1999,26:112~125.
[84] Zhu G,Chang Y,Zuo J,et al.Fudenine,a C terminal truncated rat homologue of mouse prominin,is blood glucose regulated and can up regulate the expression of GAPDH [J].Biochem Biophys Res Commun,2001,281(4): 951~956
[85] Tricarico C,Pinzani P,Bianchi S,et al.Quantitative real-time reverse transcription polymerase chain reaction: normalization to rRNA or single housekeeping genes is inappropriate for human tissue biopsies [J].Anal Biochem,2002,309(2): 293~300
[86] 陈凤花,王琳.实时荧光定量 RT-PCR内参基因的选择[J].临床检验杂志,2005,23(5): 393~395
[87] Qigui Wang,Hui Li,Ning Li.Cloning and Characterization of Chicken Adipocyte Fatty Acid Binding Protein Gene[J].ANIMAL BIOTECHNOLOGY.2004,15(2):121~132
[88] 吴建平.不同肉羊品种体脂脂肪酸遗传变异性及其特性的研究[D].甘肃:甘肃农业大学,2000
[89] 萨姆布鲁克 J,拉赛尔D W.著,黄培堂等译.分子克隆实验指南(第3版)[D].北京:科学出版社,2003
[90] SouazéF,Ntodou-ThoméA,Tran CY,RostèneW,ForgezP.QuantitativeRT-PCR:lmi its and accuracy[J]. Biotechniques,1996,21:280~285
[91] 卢缓.沙葱、地椒风味活性成分及其对绵羊瘤胃发醉和羊肉风味的影响[D].内蒙:内蒙古农业大学
[92] 包鹏云.不同品种猪肉品质和血液生化指标的比较研究:[D].内蒙:内蒙古农业大学,2000
[93] 戴瑞彤等.肉类质量的研究进展.肉类研究,2000,(2):11~13.
[94] 陈代文,张克英,胡祖禹.猪肉品质的形成原理[J].四川农业大学学报,2002,1:60～66.
[95] 林 玲.定量PCR技术的研究进展[J].国外医学遗传学分册,1999,22(3):116~120.
[96] 高海波,张拴林,陈燕红.半定量RT-PCR在基因表达方面的应用[J].畜禽业.2008,2(226):34~37.
[2] 陈润生.优质猪肉的指标及其度量方法[J].东北农业大学学报.
[3] 吴 伟,孙飞舟,胡勇军.百日龄羔羊肉肉质分析测定[J].中国草食动物,2000,2(2):32~33.
[4] 陈代文,张克英,胡祖禹.猪肉品质特征的形成原理[J].四川农业大学学报,2002,20(1): 60~66.
[5] 胡宝利.不同年龄秦川牛胴体性状与肉质性状的研究[D].陕西:西北农业大学,2001
[6] Brennand,C.P.Ha J.K.and Lindsay,R.C.Aroma Properties and Thresholds of Some Branched-chain and otherminor volatile fatty acid soccurring in milk fat meat lipids[J].J.Sensory Stud.1989,4:105~120
[7] Vignal A,Milan D,San Cristobal M, et al.A review on SNP and other types of molecular markers and their use in animal genetics[J].Genetics Selection Evolution,2002’34:275~305.
[8] 王悦生等。金华猪肉品质的研究[J].浙江农业大学学报,1982.8.
[9] WilliamsJL.The impact of marker-assisted selection in animal breeding and biotechnology [J].Scientific and Technical Review,2005,24:379~391.
[10] Womack J E.The impact of sequencing the bovine genome[J].Australian Journal of Experimental Agriculture,2006,46(2):151~153.
[11] Anderson H J.What is pork quality[A].Inc:Quality of meat and fat in pigs as affected by genetic and nutrition.
[12] 张伟力.现代猪肉品质概念与检测手段进展[J].养猪,2004,5:45～47
[13] Roschnik R.New trends in consumer acceptant and convenient[A].In:Proceedings of EuroFood ChemV,Ver sailles[C].France,1989,30～39.
[14] Novakofski J.Adipogenesis: Usefulness of in vitro and in vivo experiment models.Journal of Animal Science,2004,82: 905~915.
[15] Hodgson R R,Davis G W,Smith G C,Savell J W,Cross H R.Relationships between pork loin palatability traits and physical characteristics of cooked chops.Journal of Animal Science,1991,69:4858~4865.
[16] Wang Y H,Byrne K A,Reverter A,Harper G S,Taniguchi M,McWilliam S M,Mannen H,Oyama K,Lehnert S A.Transcriptional profiling of skeletal muscle tissue from two breeds of cattle.Mammalian Genome,2005,16: 201~210.
[17] 李凤娜,王继成,陈晓安.猪肉品质及其影响因素的研究概况[J].畜禽业.2006,6(199):12~15.
[18] Monin G,Ouali A.Muscle dierentiation and meat quality [M].InR Lawrie (Ed.): Developments in meat science (5thed),1991,(pp.89~157).
[19] Karlsson A,Enfalt A C,Essen-Gustavsson B,et al.Muscle histochemical and biochemical properties in relation to meat quality during selection for increased lean tissue growth rate in pigs [J].J Anim Sci,1993,71: 930~938.
[20] OHalloran G R,Troy D J,Buckley D J,et al.The role of endogenous proteases in the tenderization of fast glycolysing muscle [J].Meat Sci,1997,47: 187~121.
[21] Renand G,Picard B,Touraille C,et al.Relationships between muscle characteristics and meat quality traits of young Charolais bulls [J].Meat Sci,2001,59: 49~60.
[22] 孙 好 学 , 田 云 , 何 宏 轩 等 . 肌 内 脂 肪 酸 影 响 因 素 和 侯 选 基 因 研 究 进 展 [J]. 动 物 医 学 进展.2006,27(11):49~53.
[23] Warriss P D.Meat Science:An Introductory Text[M].CABI Publishing,2000.
[24] LawrieR A,Meat Science[M].Pergamon Press,1991.
[25] 黄启超.猪肉脂肪概念及其测定方法研究[J].食品科技.2006,8:134~139.
[26] 徐宁迎,赵兴波,蒋思文.猪鸡肉质性状分子标记及主效基因的研究进展[J].中国畜牧杂志,2004,4: 42~44.
[27] Farmer L J.The role of nutrients in meat ?avour formation [J].Proceedings of the Nutrition Society,1994,53: 327~333
[28] Liu Xiao-yan.Studies on Polymorphisms of H–FABP Gene and the Association With Intra muscular Fat Content in Pigs[D]
[29] HUANG Zhi-Guo,XIONG Li,LIU Zhen-Shan,The Developmental Changes and Effect on IMF Content of H-FABP and PPARγ mRNA Expression in Sheep Muscle[J].Acta Genetica Sinica,June 2006,33 (6):507~514.
[30] Williams J L.The use of marker-assisted selection in animal breeding and biotechnology [J].Scientific and Technical Review,2005,24: 379~391.
[31] wustone456.什么是候选基因.http://zhidao.baidu.com/question/41361388.html.2007-12-10 17:01
[32] Vignal A,Milan D,San Cristobal M,et al.A review on SNP and other types of molecular markers and their use in animal genetics [J].Genetics Selection Evolution,2002,34: 275~305.
[33] Veerkamp J H,van Moerkerk H B T,Prinsen C F M,et al.Structural and functional study on different human FABP types [J].Molecular and Cellular Biochemistry,1999,192: 137~142.
[34] RNA提取和RT-PCR.http//www.biorule.com,service@biorule.com.2007-4-17 20:33:34.
[35] Ockner RK.Histone overview of studies on fatty acid-binding proteins[J].Mol Cell Biochem.1990,98(1-2):3~9.
[36] Reese-Wagoner A,Thompson J,Banaszak L.Structural properties of the adipocyte lipid binding protein [J].Biochimica et Biophysica Acta,1999,1441: 106~116.
[37] Gerbens F,Jansen A,van Erp AJ,et al.The adipocyte fatty acid-binding protein locus:characterization and association with intramuscular fat content in pigs[J].Mamm Genome.1998,9(12):1022~1026
[38] Likic V A,Juranic N,Macuea S,et al.A structural water molecule in the family of fatty acid binding proteins [J].Prot Sci,2000,9: 497~504
[39] óvilo C,Pérez-Enciso M,Barragán C,et al.A QTL for intramuscular fat and backfat thickness is located on porcine chromosome 6 [J].Mamm Genome,2000,11: 344–346
[40] 叶满红.鸡脂肪酸结合蛋白基因的克隆及其与肌内脂肪含量的关系[D].2003,6
[41] Fischer H,Gustafsson T,Sundberg C J,et al.Fatty acid binding protein 4 in human skeletal muscle [J].BBRC,2006,346: 125~130
[42] 张 桂 香 ,9 个 猪 种 H-FABP 基 因 5’- 上 游 区 和 第 二 内 含 子 的 遗 传 变 异 [J]. 畜 牧 兽 医 学报,2002,33(4):304~343
[43] 李 桢 ; 曹 红 鹤 ; 储 明 星 . 中 外 11 个 猪 种 H-FABP 基 因 PCR-RFLP 的 研 究 [J]. 畜 牧 兽 医 学报,2003,34(4),313~317.
[44] Nechtellberger D,Pires V,Soolknet J,et al.Intramuscular fat content and genetic variants at fatty acid-binding protein loci in Austrian pigs[J].Anim Sci,2001,79(11):2798~2804.
[45] 陈敏.DHEA、运动通过PPARα对大鼠肝脏、心脏及肌肉中脂肪分解的调节作用[D].
[46] Lemberger T ,B Desvergne ,W Wahli .Peroxisome proliferator-activated receptors : A nuclear receptor signaling pathway in lipid physiology[J].Annu.Rey.Cell Dev.Biol,1996 ,12 :335～363.
[47] Schoonjans K,B Staels ,J Auwerx ,Role of the peroxisome proliferator-activated receptor (PPAR) in mediating the effects of fibrates and fatty acids on gene expression[J].Lip.Res.,1996 ,37 :907～925.
[48] Isabelle Issemann ,Stephen Green.Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators[J].Nature ,1990,347:645～650.
[49] 孟和,李辉,王宇祥.鸡PPARs基因组织表达特性的研究[J].遗传学报.2004,31(7):682~687.
[50] H.Sundvold,A.Brzozowska,and S.Lien. Characterisation of Bovine Peroxisome Proliferator-Activated Receptorsyγ1 and γ2:Genetic Mapping and Diferential Expression of the Two isoforms [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,1997,239,857-861.
[51] Mangelsdorf DJ,Thrmmel C,Beato M,et a1.The nuclear receptor superfamily:the second decade.Cell 1995;83:835-83.
[52] 张乾勇.PPAR的结构与功能及其生物学作用[J].国外医学卫生学分册,2000;27(5):284~288.
[53] Elangbam C.S,Tyler R D,Lightfoot RM.Peroxisome proliferator-activated receptors in atherosclerosis and inflammation an update[J].Toxicol Patho1 2001;29(2):224~231.
[54] Braissant O,Foufelle F,Scoto C,et a1.Differential expression of peroxisome proliferator-activated receptors(PPARs):tissue distribution of PPAR-alpha,-beta,and -gamma in the adult rat.Endocrinology 1996;137:354-366.
[55] Mattew E ,Poynter, Daynes R ATeroxisome proliferator-activated receptor a activation modulates cellular redox status,represses nuclear factor-kB signaling,and reduces inflammatory cytokine production in aging.J Biol Chem 1998;273(49):32833-32841
[56] Moras D, Gronemeyer H. The nuclear receptor ligand binding domain:st ructure and function. CurrO pin Cell Biol,1998 ,10 :381391
[57] Devchand PR,Keller H,Peters JM,et al.The PPAR a leukotriene B4 pathway to inflammation control.Nature 1996;384:39-43
[58] Martin NF ,Furman C ,D ele riv e Pet a1.Peroxisome proliferator-activated receptor activators inhibit oxidized low-density lipoprotein-induced endothelin-1 secretion in endothelial cells.J Cardiovasc Pharmacol 2002;40(6):822-831
[59] Elangbam C S, Tyler R D, Light foot RM.Peroxisome proliferator-activated receptors in atherosclerosis and inflammation an update.ToxicolPa tho12001;29(2):224-231.
[60] Stacls.B ,Wolfgang K, Habib A, et a1.Activation of human aortic smooth-muscle cells is inhibited by PPAR a but not PPARγactivators. Nature 1998;393:790-793
[61] Fajias L,Auboeuf D.The organization,promoter analysis,and expression of the human PPARγgene[J].The Journal of Bio-logical Chemistry,1997,272(30):18779-18789.
[62] 孟和 , 王桂华 , 王启贵 . 鸡 PPAR 基因单核苷酸多态与脂肪性状相关的研 究 [J]. 遗传学报,2002,29(2):119~123.
[63] Tontonoz P E Hu,B M Spoegelman.Stimulation of adipogenesis in fibroblasts by PPARγ2,a lipid-activated transcription factor[J].Cell,1994,79:1147~1156.
[64] 罗敏,左祥生.核转录因子PPARγ与肥胖、糖尿病的临床联系及进展[J].辽宁实用糖尿病杂志2001,9 (2):49~51Dreyer C H Keller,A Mahfoudi,V Laudet,G Krey,W Wahli Positive,Regulation of theperoxisomal betaoxidation pathway by fatty acids through activation of peroxisomal proliferator-activated receptors(PPAR)[J].Biol.Cell,1993,77:67~76
[65] Hertzel A V,Bennaars-Eiden A,Bernlohr D A.Increased lipolysis in transgenic animals over expressing the epithelial fatty acid binding protein in adipose cell [J].Lipid Res,2002,43(12): 2105~2111.
[66] Lim H,Gupta R,Ma Wet al.Cycloxygenase-2 derived prostacyclin mediates embryo implantation in the mouse via PPARδ[J].Genes Dev1999;13:1561一1574.
[67] Foman BM, Tontonoz P, Chen J, et a1. 15-Deoxy-12,14 prostaglandin J2 is a ligand for the adipocyte determination Prostaglandin factor PPAR[J].Cell,19 95,83 :80 3-812.
[68] Issemann.L .and Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators[J].Nature,1990,34 7(6295):64 5-650
[69] HUANG Zhi-Guo,XIE Zhuang.The research development of livestock and poultry adipose related gene[J].Animal Husbandry & Veterinary Medicine,2004,36(4):41~43.
[70] C Diot ,MDuaire.Characterization of a cDNA sequence encoding the peroxisome proliferator- activated receptor α in the chicken[J].Poultry Science.1999,78:1198～1202.
[71] Evans R M.The steroid and thyroid hormone receptor superfamily [J ].Sc ience,19 88,24 0:88 9-895.
[72] Chawla A, Schwarz E J, et al. Peroxisome proliferator-activated receptor (PPAR) gamma:adi pos e-p re dom inantex pressionan din ductione arlyin a dipocyted iferentiation[J ].En docrinology,199 4, 13 5( 2): 79 8-800.
[73] 姜俊芳.脂肪细胞分化相关因子基因表达在猪生长过程中的变化规律研究[D] 杭州:浙江大学,2006
[74] Xu HE,Lambert MH,Valcric G,et al.Moleecular recognition of fatty acidsby perocisome proliferators-activated receptors[J].Mo Cell,199,3(3):397-403.
[75] RNA 提取策略及RT-PCR技术.http//www.biorule.com,service@biorule.com
[76] Bligh,E,G,.Dyer,W,J.A rapid method of total lipid extraction and purification and purification[j].Can.Biochem Physiol.1959(37):911~917
[77] May J,Costa M,Ojeda S R.Developmental expression of the genes encoding transforming growth factor alpha and its receptor in the by prothalamus of female thesus macaques[J].Neurcendocrino, 1994, 60(1):346.
[78] Stanley K K,Szewczuk E.Multiplexed tandem PCR: gene profiling from small amounts of RNA using SYBR Green detection [J].Nucleic Acids Research,2005,33(20): e180
[79] 徐春兰,汪以真.半定量RT-PCR法分析猪肌肉组织细胞谷胱甘肽过氧化物酶mRNA表达水平[J].中国兽药杂志.2005,39(8):3~6;23.
[80] 陈昕,王保莉,曲东,等.小麦硫转运蛋白基因半定量RT-PCR检测方法的建立[J].西北植物学报,2006,26(2):0309~0313.
[81] Chen D,Klebe R J.Controls for validation forrelative reverse transcription-polymerase chain reaction assays.PCR Methods Appl,1993,3:127~129.
[82] Marone M,scambia G,Giannitelli C,et al.Cyclin E and cdk2 alterations in ovarian cancer: Amplification and overexpression .Int J Cancer,1997,74:390~395
[83] FreemanW M,WalkerS J,VranaK E.Quantitative RT-PCR:Pitfalls and potential[ J].BioTechniaues, 1999,26:112~125.
[84] Zhu G,Chang Y,Zuo J,et al.Fudenine,a C terminal truncated rat homologue of mouse prominin,is blood glucose regulated and can up regulate the expression of GAPDH [J].Biochem Biophys Res Commun,2001,281(4): 951~956
[85] Tricarico C,Pinzani P,Bianchi S,et al.Quantitative real-time reverse transcription polymerase chain reaction: normalization to rRNA or single housekeeping genes is inappropriate for human tissue biopsies [J].Anal Biochem,2002,309(2): 293~300
[86] 陈凤花,王琳.实时荧光定量 RT-PCR内参基因的选择[J].临床检验杂志,2005,23(5): 393~395
[87] Qigui Wang,Hui Li,Ning Li.Cloning and Characterization of Chicken Adipocyte Fatty Acid Binding Protein Gene[J].ANIMAL BIOTECHNOLOGY.2004,15(2):121~132
[88] 吴建平.不同肉羊品种体脂脂肪酸遗传变异性及其特性的研究[D].甘肃:甘肃农业大学,2000
[89] 萨姆布鲁克 J,拉赛尔D W.著,黄培堂等译.分子克隆实验指南(第3版)[D].北京:科学出版社,2003
[90] SouazéF,Ntodou-ThoméA,Tran CY,RostèneW,ForgezP.QuantitativeRT-PCR:lmi its and accuracy[J]. Biotechniques,1996,21:280~285
[91] 卢缓.沙葱、地椒风味活性成分及其对绵羊瘤胃发醉和羊肉风味的影响[D].内蒙:内蒙古农业大学
[92] 包鹏云.不同品种猪肉品质和血液生化指标的比较研究:[D].内蒙:内蒙古农业大学,2000
[93] 戴瑞彤等.肉类质量的研究进展.肉类研究,2000,(2):11~13.
[94] 陈代文,张克英,胡祖禹.猪肉品质的形成原理[J].四川农业大学学报,2002,1:60～66.
[95] 林 玲.定量PCR技术的研究进展[J].国外医学遗传学分册,1999,22(3):116~120.
[96] 高海波,张拴林,陈燕红.半定量RT-PCR在基因表达方面的应用[J].畜禽业.2008,2(226):34~37.