鹅黑素皮质素受体-4基因（MC4R）单核苷酸多态性分析与生产性状相关研究
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摘要
黑素皮质素受体家族包括MC1R、MC2R、MC3R、MC4R和MC5R,是目前所知的最小的G蛋白偶联受体亚家族,它们均属于A类7个跨膜α螺旋G蛋白受体,是一系列小基因的产物。作为其中的家族成员基因,MC4R基因由332个氨基酸残基组成,具有七次跨膜结构,在肾上腺、性腺、脾、脂肪和脑等组织表达,MC4R基因具有介导Leptin蛋白的功能,是一个调节能量与能量动态平衡的重要的信号分子,其主要作用是控制食欲、体重、能量代谢。
本研究采用PCR-SSCP和DNA测序的方法,对朗德鹅、四川白鹅和皖西白鹅三个品种共175个个体进行了MC4R基因外显子多态性的研究,结果表明,MC4R基因外显子存在M、N两个多态位点,分别位于G108 A、C 627 T,并且在各个品种中都存在多态性,两个SNP位点均为同义突变。χ~2适合性检验表明:位点M、N已经达到Hardy-Weinberg平衡状态(P＞0.05)。
研究同时对朗德鹅94个个体进行相关分析,各位点基因型与屠体性状关联分析得知:M位点各基因型与屠体重(CW)、胸肌率(BMP)和腿肌率(LMP)有显著(P＜0.05)影响,N位点各基因型与活重(BW)、屠体重(CW)、半净膛重(SEW)和全净膛重(EW)有显著(P＜0.05)影响。与产肝性状关联分析得知:N位点各基因型与肝体比(LBR)和肝屠比(LCR)显著(P＜0.05)影响。两位点对鹅的脂肪沉积性能均无显著影响,三种基因型效应差异都不显著(P＞0.05)。
研究还通过对各复合位点与朗德鹅屠宰性状、产肝性状和脂肪沉积性状进行相关分析,发现H3H3基因型具有最高的活重(BW)、屠体重(CW)、半净膛重(SEW)和胸肌重(BMW),与其他单倍型组合差异性显著;H4H4基因型具有最高的肝重(LW)、肝体比(LBR)、肝屠比(LCR)和腹脂率(AFR)与其他单倍型组合差异性显著,说明H3H3和H4H4型是最佳基因型组合。本实验初步推断MC4R基因可能是影响鹅屠体性状的主效基因或与主效基因连锁,且N位点可能是影响鹅MC4R基因产肝性状的主要位点。
Melanocortin Receptors family(MCRs,including MC1R、MC2R、MC3R、MC4R and MC5R) is the smallest G-protein-coupled receptor subfamily.All of the members are the productions of a series small genes which belong to species Aα-Helix G-protein receptor including 7 transmembrane domains.As one of MCRs,MC4R gene codes 332 amino acids residues constructing 7 transmembrane domains,which expresses in adrenal gland,gonad,spleen,fatty,brain and other tissues.MC4R gene is an important signaling molecule mediating leptin protein to adjust energy and energy development balance,in order to control the appetite and energy metabolism.
In this study,3 breeds(including Landes geese,Sichuan white geese and Wanxi white geese 175 individuals)were used to analyze the polymorphism of the exon of MC4R gene by PCR-SSCP and DNA sequencing.Results indicated that there were two mutation sites named M(C 108 T) and N(G 627 A),which were both samesense mutations found in each geese line.Statistical results showed that the polymorphic sites fitted with Hardy-Weinberg equilibrium for site M and N(P＞0.05).
In this study,94 Landes geese were used to statistics analysis,and the results showed that in slaughter traits,there were significant differences(P＜0.05) for carcass weight(CW),breast muscle percentage(BMP) and leg muscle percentage(LMP) in M site,and significant differences(P＜0.05) for body weight(BW),carcass weight(CW), semi-eviscerated weight(SEW) and eviscerate weight(EW) in N site.In addition in N site there were significant differences(P＜0.05) for liver and body ratio(LBR) and liver and carcass ratio(LCR) in fatty liver performance.The two sites had no effect on the fat deposition performance,which means there was no significant difference(P＞0.05) among the three genotypes.
The statistics analysis results of haplotype on carcass traits,fatty liver and fat deposition performance showed that there were the highest body weight(BW),carcass weight(CW),semi-eviscerated weight(SEW) and breast muscle weight(BMW) in diplotype H3H3,liver weight(LW) liver and body weight ratio(LBR),liver and carcass weight ratio(LCR) and abdominal fat weight(AFR) in diplotype H4H4,and there were significant differences with other genotypes,from which could be concluded that H3H3 and H4H4 genotype were the best diplotype.It could be concluded from the results that MC4R gene might be the major gene affecting the meat traits of geese or linking with the major gene,and the N site may be the major site affecting the fatty liver performance of geese.
本研究采用PCR-SSCP和DNA测序的方法,对朗德鹅、四川白鹅和皖西白鹅三个品种共175个个体进行了MC4R基因外显子多态性的研究,结果表明,MC4R基因外显子存在M、N两个多态位点,分别位于G108 A、C 627 T,并且在各个品种中都存在多态性,两个SNP位点均为同义突变。χ~2适合性检验表明:位点M、N已经达到Hardy-Weinberg平衡状态(P＞0.05)。
研究同时对朗德鹅94个个体进行相关分析,各位点基因型与屠体性状关联分析得知:M位点各基因型与屠体重(CW)、胸肌率(BMP)和腿肌率(LMP)有显著(P＜0.05)影响,N位点各基因型与活重(BW)、屠体重(CW)、半净膛重(SEW)和全净膛重(EW)有显著(P＜0.05)影响。与产肝性状关联分析得知:N位点各基因型与肝体比(LBR)和肝屠比(LCR)显著(P＜0.05)影响。两位点对鹅的脂肪沉积性能均无显著影响,三种基因型效应差异都不显著(P＞0.05)。
研究还通过对各复合位点与朗德鹅屠宰性状、产肝性状和脂肪沉积性状进行相关分析,发现H3H3基因型具有最高的活重(BW)、屠体重(CW)、半净膛重(SEW)和胸肌重(BMW),与其他单倍型组合差异性显著;H4H4基因型具有最高的肝重(LW)、肝体比(LBR)、肝屠比(LCR)和腹脂率(AFR)与其他单倍型组合差异性显著,说明H3H3和H4H4型是最佳基因型组合。本实验初步推断MC4R基因可能是影响鹅屠体性状的主效基因或与主效基因连锁,且N位点可能是影响鹅MC4R基因产肝性状的主要位点。
Melanocortin Receptors family(MCRs,including MC1R、MC2R、MC3R、MC4R and MC5R) is the smallest G-protein-coupled receptor subfamily.All of the members are the productions of a series small genes which belong to species Aα-Helix G-protein receptor including 7 transmembrane domains.As one of MCRs,MC4R gene codes 332 amino acids residues constructing 7 transmembrane domains,which expresses in adrenal gland,gonad,spleen,fatty,brain and other tissues.MC4R gene is an important signaling molecule mediating leptin protein to adjust energy and energy development balance,in order to control the appetite and energy metabolism.
In this study,3 breeds(including Landes geese,Sichuan white geese and Wanxi white geese 175 individuals)were used to analyze the polymorphism of the exon of MC4R gene by PCR-SSCP and DNA sequencing.Results indicated that there were two mutation sites named M(C 108 T) and N(G 627 A),which were both samesense mutations found in each geese line.Statistical results showed that the polymorphic sites fitted with Hardy-Weinberg equilibrium for site M and N(P＞0.05).
In this study,94 Landes geese were used to statistics analysis,and the results showed that in slaughter traits,there were significant differences(P＜0.05) for carcass weight(CW),breast muscle percentage(BMP) and leg muscle percentage(LMP) in M site,and significant differences(P＜0.05) for body weight(BW),carcass weight(CW), semi-eviscerated weight(SEW) and eviscerate weight(EW) in N site.In addition in N site there were significant differences(P＜0.05) for liver and body ratio(LBR) and liver and carcass ratio(LCR) in fatty liver performance.The two sites had no effect on the fat deposition performance,which means there was no significant difference(P＞0.05) among the three genotypes.
The statistics analysis results of haplotype on carcass traits,fatty liver and fat deposition performance showed that there were the highest body weight(BW),carcass weight(CW),semi-eviscerated weight(SEW) and breast muscle weight(BMW) in diplotype H3H3,liver weight(LW) liver and body weight ratio(LBR),liver and carcass weight ratio(LCR) and abdominal fat weight(AFR) in diplotype H4H4,and there were significant differences with other genotypes,from which could be concluded that H3H3 and H4H4 genotype were the best diplotype.It could be concluded from the results that MC4R gene might be the major gene affecting the meat traits of geese or linking with the major gene,and the N site may be the major site affecting the fatty liver performance of geese.
引文
[1]顾春云.鹅的生物学特性及饲养管理技术[J].养禽与禽病防治,2002,11(1):17-17.
[2]刘五岳.鹅与养鹅技术[J].当代畜禽养殖业,2005(7):34-35.
[3]闵育娜,高玉鹏,候水生,等.我国养鹅业现状及前景展望[J].中国畜牧杂志,2005,41(5):57-59.
[4]黄治国,谢庄.畜禽脂肪性状相关基因研究进展[J].畜牧与兽医,2004,36(4):41-43.
[5]Tatro J B.Melanotropin receptors in the brain are differentially distributed and recognize both corticotrophin and α-melanocyte stimulating hormone[J].Brain Research,1990,536(1):124-132.
[6]Chhajlani V,Wikberg JE.Molecular cloning and expression of the human melanocyte stimulating hormone receptor cDNA[J].FEBS lett,1992,309(3):417-420.
[7]Makova K,Norton H.Wetldwide polymorphism at the MC1R locus and normal pigmentation variation in humans[J].Peptides,2005(26):1901-1908.
[8]Takeuchi S,Takahashi S.A possible involvement of melanocortin-3 receptor in the regulation of adrenal gland function in the chicken[J].Biochim Biophys Acta,1999,1448(3):512-518.
[9]蒋思文,彭键,熊远.黑素皮质素受体对动物采食量和能量稳态的调控[J].遗传,2002,24(2):223-226.
[10]Mountjoy KG,Mortrud MT,MJ Low,et al.Localization of the melanocortin-4 receptor(MC4R)in neuroendocrine and autonomic control circuits in the brain[J].Mol Endocfinol,1994,8(1):1298-1308.
[11]Schioth HB,Muceniece R,Larsson M,et al.The melanocortin 1,3,4 or 5 receptors do not have a binding epitope for ACTH beyond the sequence of alpha-MSH[J].J Endocrinol,1997,155(1):73-78.
[12]Lagerstrom MC,Klovins J,Fredriksson R,el al.High affinity agonistic metal ion binding sites within the melanocortin-4 receptor illustrate conformational change of TM3[J].JBC,2003,278(51):51521-51526.
[13]Srinivasan S.,Lubrano-Berthelier C.,Govaerts,et al.Constitutive activity of the melanocortin-4receptor is maintained by its N-terminal domain and plays a role in energy homeostasis in humans[J].Journal of Clinical Inverstigation,2004,10,114(8):1158-1164.
[14]Gantz I,Shimoto Y,J Chambers,et al.Molecular cloning,expression,and characterization of a fifth melanocortin receptor[J].Biochem B iophys Rescommun,1994,200(3):1214-1220.
[15]Haegeman A,Coopman F,Jacobs K,et al.Bovine melanocortin receptor 4:cDNA sequence,polymerphisms and mapping[J].AnimGenet,2001,32(4):189-192.
[16]Lubrano-Bertbelier C,Cavazos M,Le Stunf C,et al.The human MC4R promoter:characterization and role in obesity[J].Diabetes,2003,52(12):2996-3000.
[17]Sinha P S,Schioth H B,Tatro J B.Roles of melanocortin-4 receptor in antipyretic and hyperthemic actions of centrally administered alpha-MSH[J].Brain Res,2004,1001(1-2):150-158.
[18]Andersson L.Melanocortin receptor variants with phenotypic effects in horse,pig and chicken [J].Ann N Y Acad Sci,2003,994(20):313-318.
[19]Hoggard N,Hunter L,Duncan J S,et al.Regulation of adipose tissue leptin secretion by alpha-MSH and agouti-related protein:Further evidence of an interaction between leptin and the melanocortin signaling system[J].J Mol Endocrinol,2004,32(1):145-153.
[20]Huszar D,Lynch CA,Fairchild-Huntress V,et al.Targeted disruption of the melanocortin 4-receptor results in obesity in mice[J].Cell,1997,88:131-141.
[21]K SKim,N J Larsen,M F Rothschild.Rapid communication:Linkage and physical mapping of the porcine melanocortin-4 receptor(MC4R) gene[J].Journal of Animal Science,2000,78(3):791-792.
[22]Sinha PS,Sehioth HB,Tatro JB,et al.Activation of central melanocortin-4 receptor suppresses lipopoly- saccharide-induced fever in rats[J].Am J Physiol Regul Integr Comp Physiol,2003,284(6):1595-1603.
[23]Farooqi IS,Yeo GS,Keogh JM,et al.Dominant and recessive inheritance of morbid obesity associated with melanocortin 4 receptor deficiency[J].J Clin Invest,2000,106(2):271-279.
[24]Siracusa LD.The agouti gene:turned on to yellow[J].Trends Genet,1994,10(5):423-428.
[25]Mihenberger R J,Mynatt RL,Wilkinson JE,et al.The role of the agouti gene in the yellow obese syndrome[J].J Nutr,1997,127(23):1902-1907.
[26]Tritos NA,Maratos Flier E.Two important systems in energy homeostasis:melanocortins and melanin concentrating hormone[J].Neuropoptides,1999,33(5):339-349.
[27]Carrie,Monck E K.Agouti-related protein functions as an inverse agonist at a constitutively active brain melanocortin-4 receptor[J].Peptides,2001,99(1):1-7.
[28]David E Cummings,Michael W.Schwartz.Melanocortins and body weight:a tale of two receptors[J].Nature Genetics,2000,26(1):8-9.
[29]Jeffrey M.Friendman.The alphabet of weight control[J].Nature,1997,385(1):119-120.
[30]Brocke KS,Neu-Yilik G,Gehring NH,et al.The human intronless melanocortin 4-receptor gene is NMD insensitive[J].Hum Mol Genet,2002,11(3):331-335.
[31]Haskell-Luevano C,Monck EK.Agouti-related protein functions as an inverse agonist at a constitutively active brain melanocortin-4 receptor[J].Regul Peptides,2001,99(1):1-7.
[32]Yeo GSH,Farooqi I S,Aminian S.A frameshift mutation in MC4R associated with dominantly inherited human obesity[J].Nature Genetics,1998,20(10):111-112.
[33]Vaisse C,Clement K,Durand E,et al.Melanocortin 4 receptor mutations are a frequent and heterogeneouscause of morbid obesity[J].Clin Invest,2000,106(1):253- 262.
[34]Farooqi I S,Yeo GSH,Keogh JM,et al.Dominant and recessive inheritance of morbid obesity associated with melanocortin 4 receptor deficiency[J].Clin Invest,2000,106(1):271-279.
[35]Sina Mani,Hinney Anke,Ziegler A,et al.Phenotypes in three pedigrees with autosomal dominant obesity caused by haploinsufficiency mutations in the MC4R gene[J].American journal of Human Genetics,1999,65(6):1502-1507.
[36]张崇本,吴鹤龄.人类单基因肥胖的遗传学基础[J].遗传学报,2004,31(8):864-869.
[37]Eun Jin Lee,Jin-Won Jung.Differential regulation of camp-mediated gene transcription and ligand selectivity by MC3R and MC4R melancortin receptors[J].Eur.J.Biochem.,2001,268(1):582-591.
[38]Huszar D,Lynch C A,Fairchild-Huntress V,et al.Targeted disruption of the melanocortin-4receptor results in obesity in mice[J].Cell,1997,88(1):131-141.
[39]Kim KS,Larsen N,Short T,et al.A missense variant of the porcine melanocortin-4 receptor gene is associatd with fatness,growth,and feed intake traits[J].Mamma Genome,2000,11(1):131-135.
[40]刘桂兰,蒋思文,熊远著,等.猪资源家系MC4R基因扫描及其与脂肪性状的相关性分析[J].遗传学报,2002,29(6):497-501.
[41]李长龙,潘玉春,孟和,等.H-FABP、MC4R、ADDl基因多态性在3个猪群中分布及其与肌内脂 肪和背膘的相关研究[J].遗传,2006,28(2):159-164.
[42]张轶博,巴彩凤,苏玉虹,等.比格犬MC4R基因多态性与体重相关性的研究[J].遗传,2006,28(10):1224-1228.
[43]蒋美山,陈仕毅,赖松家,等.兔黑素皮质素受体4(MC4R)基因多态性及其与体重及屠体性状的关联研究[J].遗传,2008,30(12):1574-1578.
[44]霍明东,王守志,李辉.MC4R基因多态性与鸡生长和体组成性状的相关研究[J].东北农业大学学报,2006,37(2):184-189.
[45]仇雪梅,李宁,邓学梅,等.鸡MC4R基因的SNPs及其与屠体性状的相关研究[J].中国科学,2006,36(2):127-133.
[46]李国辉,王金玉,陶勇,等.京海黄鸡MC4R基因新突变位点的分析[J].畜牧兽医杂志,2008,27(2):28-30.
[47]张同燕,陈宏权,梁忠,等.鹅黑素皮质素受体-4基因的克隆与序列分析[J].畜牧兽医学报,2007,38(9):913-919.
[47]Beckmann J.S,Soller M.Toward a unified approach to genetic mapping of eukaryotes based on sequence tagged microsatellite sites[J].Biotechnology,1990,8(10):930-932.
[48]苏毅.优质肉鸡黑素皮质素3、4基因单核苷酸多态性与生产性状相关研究[D].雅安:四川农业大学,2006:60-65.
[49]顿桂玲.猪黑素皮质素受体1(MC1R)基因与毛色表型相关及5`UTR变异研究[D].石家庄:河北农业大学,2007:50-56.
[50]王琼.MyoD,MyoG基因的多态性与鸡屠宰性状和肉质性状的相关性研究[D].雅安:四川农业大学,2007:55-58.
[51]Orita M,Lwahana H,Kanazawa H,et al.Detection of polymorphisms of human DNA by gel electrophore- sisas singles trand conformation polymorphisms[J].Proc Natl Acad Sci USA,1989,86(8):2766-2770.
[52]Orita M,Sekiya T,Hayashi K.DNA sequence Polymophism in Alurepeats[J].Genomics,1998,8(2):271-278
[53]Hayashi K.A single and sensitive method for detection of mutations in the genomic DNA.PCR method and Applications,PCR-SSCP,1992,(1):34-38.
[54]欧阳建华,黄建安.PCR-SSCP技术的研究进展[J].上海畜牧兽医通讯,2002(4):10-11.
[55]孙万元.长金猪及其肉质性状候选基因的研究[D].杭州:浙江大学,2004:16-18.
[56]沐建煜,周伟.朗德鹅肥肝生产技术[J].郑州牧业工程高等专科学校学报,2005,8(1):174-175.
[57]苏世广,张朝霞,许月英,等.皖西自鹅不同饲养方式采食量及生长速度测定[J].安徽农业科学.2001,29(3):398-409.
[58]卢圣栋.现代分子生物学实验技术(第2版)[M].北京:中国协和医科大学出版社,1999:175-180.
[59]李建华.四川白鹅的品种特性及杂交应用[J].中国家禽,2003,25(10):34-36.
[60]储明星译.师守堃 校.数量遗传学导论[M].北京:中国农业科技出版社.2000:57-60.
[61]吴泽辉.生长分化因子9基因多态性与济宁青山羊产羔数关系的研究[D].四川农业大学,2005:31-32.
[62]李春喜,王志和,王文林.生物统计学[M].北京:科学出版社,2003:154-156.
[63]明道绪.高级生物统计与实验设计[M].中国农业出版社,2000:210-212.
[64]吴登俊.SAS高级统计分析系统[M].雅安:四川农业大学,2002:240-245.
[65]Stephens M,Smith N,Donnelly P.A new statistical method for haplotype reconstruction from population data[J].American Journal Of Human Genetics,2001,68(1):978-989.
[66]黄艳群,陈文,邓雪梅,等.鸡lmbrl基因外显子16的SNP检测和单倍型分析[J].畜牧兽医学报,2007,38(8):781-785.
[67]Fukuoko Y,Kuhl D.P.A.,Pizzuti A.et al.Variation of the CGG repeat at the fragile X site results in genetic instability:resolution of the sherman paradox[J].Cell,1992,67(1):1047-1058.
[68]姜远良,李宁.影响PCR-SSCP的因素分析[J].农业生物技术学报,2000,8(3):245-247.
[69]刘佳建,李奎,彭中镇.甘油浓度对PCR-SSCP分辨效果的影响[J].生物技术通讯,1998,2(1):160-161.
[70]Takeuchi S,Takahashi S.Melanocortin receptor genes in the chicken-Tissue distributions[J].Gen Comp Endocrinol,1998,112(2):220-231.
[71]Haegeman A,Coopman F,Jacobs K,et al.Bovine melanocortin receptor 4:cDNA sequence,polymorphisms and mapping[J].AnimGenet,2001,32(4):189-192.
[72]杨秀芹,于浩,顾志刚,等.野猪MC4R基因的克隆及变异初步研究[J].动物学杂志,2007,42(2):71-74.
[73]陈宏权,黄华云,陈华,等,鹅MC4R基因RFLP及其胴体和羽绒性状的关联性[J].畜牧兽医学报,2008,39(7):885-890.
[74]孙乃恩,孙东旭,朱德熙.分子遗传学(第1版)[M].南京:南京大学出版社,1990:249-257.
[2]刘五岳.鹅与养鹅技术[J].当代畜禽养殖业,2005(7):34-35.
[3]闵育娜,高玉鹏,候水生,等.我国养鹅业现状及前景展望[J].中国畜牧杂志,2005,41(5):57-59.
[4]黄治国,谢庄.畜禽脂肪性状相关基因研究进展[J].畜牧与兽医,2004,36(4):41-43.
[5]Tatro J B.Melanotropin receptors in the brain are differentially distributed and recognize both corticotrophin and α-melanocyte stimulating hormone[J].Brain Research,1990,536(1):124-132.
[6]Chhajlani V,Wikberg JE.Molecular cloning and expression of the human melanocyte stimulating hormone receptor cDNA[J].FEBS lett,1992,309(3):417-420.
[7]Makova K,Norton H.Wetldwide polymorphism at the MC1R locus and normal pigmentation variation in humans[J].Peptides,2005(26):1901-1908.
[8]Takeuchi S,Takahashi S.A possible involvement of melanocortin-3 receptor in the regulation of adrenal gland function in the chicken[J].Biochim Biophys Acta,1999,1448(3):512-518.
[9]蒋思文,彭键,熊远.黑素皮质素受体对动物采食量和能量稳态的调控[J].遗传,2002,24(2):223-226.
[10]Mountjoy KG,Mortrud MT,MJ Low,et al.Localization of the melanocortin-4 receptor(MC4R)in neuroendocrine and autonomic control circuits in the brain[J].Mol Endocfinol,1994,8(1):1298-1308.
[11]Schioth HB,Muceniece R,Larsson M,et al.The melanocortin 1,3,4 or 5 receptors do not have a binding epitope for ACTH beyond the sequence of alpha-MSH[J].J Endocrinol,1997,155(1):73-78.
[12]Lagerstrom MC,Klovins J,Fredriksson R,el al.High affinity agonistic metal ion binding sites within the melanocortin-4 receptor illustrate conformational change of TM3[J].JBC,2003,278(51):51521-51526.
[13]Srinivasan S.,Lubrano-Berthelier C.,Govaerts,et al.Constitutive activity of the melanocortin-4receptor is maintained by its N-terminal domain and plays a role in energy homeostasis in humans[J].Journal of Clinical Inverstigation,2004,10,114(8):1158-1164.
[14]Gantz I,Shimoto Y,J Chambers,et al.Molecular cloning,expression,and characterization of a fifth melanocortin receptor[J].Biochem B iophys Rescommun,1994,200(3):1214-1220.
[15]Haegeman A,Coopman F,Jacobs K,et al.Bovine melanocortin receptor 4:cDNA sequence,polymerphisms and mapping[J].AnimGenet,2001,32(4):189-192.
[16]Lubrano-Bertbelier C,Cavazos M,Le Stunf C,et al.The human MC4R promoter:characterization and role in obesity[J].Diabetes,2003,52(12):2996-3000.
[17]Sinha P S,Schioth H B,Tatro J B.Roles of melanocortin-4 receptor in antipyretic and hyperthemic actions of centrally administered alpha-MSH[J].Brain Res,2004,1001(1-2):150-158.
[18]Andersson L.Melanocortin receptor variants with phenotypic effects in horse,pig and chicken [J].Ann N Y Acad Sci,2003,994(20):313-318.
[19]Hoggard N,Hunter L,Duncan J S,et al.Regulation of adipose tissue leptin secretion by alpha-MSH and agouti-related protein:Further evidence of an interaction between leptin and the melanocortin signaling system[J].J Mol Endocrinol,2004,32(1):145-153.
[20]Huszar D,Lynch CA,Fairchild-Huntress V,et al.Targeted disruption of the melanocortin 4-receptor results in obesity in mice[J].Cell,1997,88:131-141.
[21]K SKim,N J Larsen,M F Rothschild.Rapid communication:Linkage and physical mapping of the porcine melanocortin-4 receptor(MC4R) gene[J].Journal of Animal Science,2000,78(3):791-792.
[22]Sinha PS,Sehioth HB,Tatro JB,et al.Activation of central melanocortin-4 receptor suppresses lipopoly- saccharide-induced fever in rats[J].Am J Physiol Regul Integr Comp Physiol,2003,284(6):1595-1603.
[23]Farooqi IS,Yeo GS,Keogh JM,et al.Dominant and recessive inheritance of morbid obesity associated with melanocortin 4 receptor deficiency[J].J Clin Invest,2000,106(2):271-279.
[24]Siracusa LD.The agouti gene:turned on to yellow[J].Trends Genet,1994,10(5):423-428.
[25]Mihenberger R J,Mynatt RL,Wilkinson JE,et al.The role of the agouti gene in the yellow obese syndrome[J].J Nutr,1997,127(23):1902-1907.
[26]Tritos NA,Maratos Flier E.Two important systems in energy homeostasis:melanocortins and melanin concentrating hormone[J].Neuropoptides,1999,33(5):339-349.
[27]Carrie,Monck E K.Agouti-related protein functions as an inverse agonist at a constitutively active brain melanocortin-4 receptor[J].Peptides,2001,99(1):1-7.
[28]David E Cummings,Michael W.Schwartz.Melanocortins and body weight:a tale of two receptors[J].Nature Genetics,2000,26(1):8-9.
[29]Jeffrey M.Friendman.The alphabet of weight control[J].Nature,1997,385(1):119-120.
[30]Brocke KS,Neu-Yilik G,Gehring NH,et al.The human intronless melanocortin 4-receptor gene is NMD insensitive[J].Hum Mol Genet,2002,11(3):331-335.
[31]Haskell-Luevano C,Monck EK.Agouti-related protein functions as an inverse agonist at a constitutively active brain melanocortin-4 receptor[J].Regul Peptides,2001,99(1):1-7.
[32]Yeo GSH,Farooqi I S,Aminian S.A frameshift mutation in MC4R associated with dominantly inherited human obesity[J].Nature Genetics,1998,20(10):111-112.
[33]Vaisse C,Clement K,Durand E,et al.Melanocortin 4 receptor mutations are a frequent and heterogeneouscause of morbid obesity[J].Clin Invest,2000,106(1):253- 262.
[34]Farooqi I S,Yeo GSH,Keogh JM,et al.Dominant and recessive inheritance of morbid obesity associated with melanocortin 4 receptor deficiency[J].Clin Invest,2000,106(1):271-279.
[35]Sina Mani,Hinney Anke,Ziegler A,et al.Phenotypes in three pedigrees with autosomal dominant obesity caused by haploinsufficiency mutations in the MC4R gene[J].American journal of Human Genetics,1999,65(6):1502-1507.
[36]张崇本,吴鹤龄.人类单基因肥胖的遗传学基础[J].遗传学报,2004,31(8):864-869.
[37]Eun Jin Lee,Jin-Won Jung.Differential regulation of camp-mediated gene transcription and ligand selectivity by MC3R and MC4R melancortin receptors[J].Eur.J.Biochem.,2001,268(1):582-591.
[38]Huszar D,Lynch C A,Fairchild-Huntress V,et al.Targeted disruption of the melanocortin-4receptor results in obesity in mice[J].Cell,1997,88(1):131-141.
[39]Kim KS,Larsen N,Short T,et al.A missense variant of the porcine melanocortin-4 receptor gene is associatd with fatness,growth,and feed intake traits[J].Mamma Genome,2000,11(1):131-135.
[40]刘桂兰,蒋思文,熊远著,等.猪资源家系MC4R基因扫描及其与脂肪性状的相关性分析[J].遗传学报,2002,29(6):497-501.
[41]李长龙,潘玉春,孟和,等.H-FABP、MC4R、ADDl基因多态性在3个猪群中分布及其与肌内脂 肪和背膘的相关研究[J].遗传,2006,28(2):159-164.
[42]张轶博,巴彩凤,苏玉虹,等.比格犬MC4R基因多态性与体重相关性的研究[J].遗传,2006,28(10):1224-1228.
[43]蒋美山,陈仕毅,赖松家,等.兔黑素皮质素受体4(MC4R)基因多态性及其与体重及屠体性状的关联研究[J].遗传,2008,30(12):1574-1578.
[44]霍明东,王守志,李辉.MC4R基因多态性与鸡生长和体组成性状的相关研究[J].东北农业大学学报,2006,37(2):184-189.
[45]仇雪梅,李宁,邓学梅,等.鸡MC4R基因的SNPs及其与屠体性状的相关研究[J].中国科学,2006,36(2):127-133.
[46]李国辉,王金玉,陶勇,等.京海黄鸡MC4R基因新突变位点的分析[J].畜牧兽医杂志,2008,27(2):28-30.
[47]张同燕,陈宏权,梁忠,等.鹅黑素皮质素受体-4基因的克隆与序列分析[J].畜牧兽医学报,2007,38(9):913-919.
[47]Beckmann J.S,Soller M.Toward a unified approach to genetic mapping of eukaryotes based on sequence tagged microsatellite sites[J].Biotechnology,1990,8(10):930-932.
[48]苏毅.优质肉鸡黑素皮质素3、4基因单核苷酸多态性与生产性状相关研究[D].雅安:四川农业大学,2006:60-65.
[49]顿桂玲.猪黑素皮质素受体1(MC1R)基因与毛色表型相关及5`UTR变异研究[D].石家庄:河北农业大学,2007:50-56.
[50]王琼.MyoD,MyoG基因的多态性与鸡屠宰性状和肉质性状的相关性研究[D].雅安:四川农业大学,2007:55-58.
[51]Orita M,Lwahana H,Kanazawa H,et al.Detection of polymorphisms of human DNA by gel electrophore- sisas singles trand conformation polymorphisms[J].Proc Natl Acad Sci USA,1989,86(8):2766-2770.
[52]Orita M,Sekiya T,Hayashi K.DNA sequence Polymophism in Alurepeats[J].Genomics,1998,8(2):271-278
[53]Hayashi K.A single and sensitive method for detection of mutations in the genomic DNA.PCR method and Applications,PCR-SSCP,1992,(1):34-38.
[54]欧阳建华,黄建安.PCR-SSCP技术的研究进展[J].上海畜牧兽医通讯,2002(4):10-11.
[55]孙万元.长金猪及其肉质性状候选基因的研究[D].杭州:浙江大学,2004:16-18.
[56]沐建煜,周伟.朗德鹅肥肝生产技术[J].郑州牧业工程高等专科学校学报,2005,8(1):174-175.
[57]苏世广,张朝霞,许月英,等.皖西自鹅不同饲养方式采食量及生长速度测定[J].安徽农业科学.2001,29(3):398-409.
[58]卢圣栋.现代分子生物学实验技术(第2版)[M].北京:中国协和医科大学出版社,1999:175-180.
[59]李建华.四川白鹅的品种特性及杂交应用[J].中国家禽,2003,25(10):34-36.
[60]储明星译.师守堃 校.数量遗传学导论[M].北京:中国农业科技出版社.2000:57-60.
[61]吴泽辉.生长分化因子9基因多态性与济宁青山羊产羔数关系的研究[D].四川农业大学,2005:31-32.
[62]李春喜,王志和,王文林.生物统计学[M].北京:科学出版社,2003:154-156.
[63]明道绪.高级生物统计与实验设计[M].中国农业出版社,2000:210-212.
[64]吴登俊.SAS高级统计分析系统[M].雅安:四川农业大学,2002:240-245.
[65]Stephens M,Smith N,Donnelly P.A new statistical method for haplotype reconstruction from population data[J].American Journal Of Human Genetics,2001,68(1):978-989.
[66]黄艳群,陈文,邓雪梅,等.鸡lmbrl基因外显子16的SNP检测和单倍型分析[J].畜牧兽医学报,2007,38(8):781-785.
[67]Fukuoko Y,Kuhl D.P.A.,Pizzuti A.et al.Variation of the CGG repeat at the fragile X site results in genetic instability:resolution of the sherman paradox[J].Cell,1992,67(1):1047-1058.
[68]姜远良,李宁.影响PCR-SSCP的因素分析[J].农业生物技术学报,2000,8(3):245-247.
[69]刘佳建,李奎,彭中镇.甘油浓度对PCR-SSCP分辨效果的影响[J].生物技术通讯,1998,2(1):160-161.
[70]Takeuchi S,Takahashi S.Melanocortin receptor genes in the chicken-Tissue distributions[J].Gen Comp Endocrinol,1998,112(2):220-231.
[71]Haegeman A,Coopman F,Jacobs K,et al.Bovine melanocortin receptor 4:cDNA sequence,polymorphisms and mapping[J].AnimGenet,2001,32(4):189-192.
[72]杨秀芹,于浩,顾志刚,等.野猪MC4R基因的克隆及变异初步研究[J].动物学杂志,2007,42(2):71-74.
[73]陈宏权,黄华云,陈华,等,鹅MC4R基因RFLP及其胴体和羽绒性状的关联性[J].畜牧兽医学报,2008,39(7):885-890.
[74]孙乃恩,孙东旭,朱德熙.分子遗传学(第1版)[M].南京:南京大学出版社,1990:249-257.