X染色体60149273位点在脂尾（臀）和瘦尾绵羊品种中的多态性及其基因定位

doi:10.3864/j.issn.0578-1752.2013.22.017

摘要/Abstract

摘要： 【目的】研究绵羊X染色体60149273位点在不同沉脂尾型绵羊品种中的多态性，为解析绵羊尾脂沉积性状的遗传机制提供基础数据。【方法】首先，采用PCR-SSCP分型方法检测X染色体60149273位点在不同脂尾类型绵羊品种的多态性，并借助PCR产物直接测序的方法确认各基因型相应序列，利用卡方检验的方法检测各基因型分布在不同品种中的平衡性。其次，利用生物信息方法将该SNP定位与绵羊androgen receptor（AR），并使用电子克隆的方法克隆出绵羊AR全长编码序列。同时以阿勒泰羊臀部脂肪RNA为材料，采用RT-PCR的方克隆绵羊AR第3—8外显子序列，利用生物信息学方法分析序列同源性以及序列的物种进化保守性。【结果】①PCR-SSCP结果显示，在脂尾（臀）型绵羊品种阿勒泰羊和湖羊群体中X染色体60149273位点未检出多态性，GG基因型占100%；而瘦尾（臀）型绵羊品种中国美利奴和萨福克羊群体中则出现了多态性，GG基因型分别下降至10%和21%。②利用比较基因组学首次电子克隆出该SNP所属基因AR的全长编码区，并从阿勒泰羊臀脂mRNA中成功扩增羊源AR第3—8外显子片段，测序结果与电子克隆序列完全一致。③物种同源性比对进一步显示，AR在哺乳类动物中高度保守，哺乳动物各物种间同源性高达90%；进化树分析结果显示，牛、绵羊、海豚和猪等哺乳动物亲缘较近。【结论】首次发现绵羊AR第3内含子一处SNP在脂尾（臀）与瘦尾绵羊品种中存在较大差异，该SNP将可作为一个分子标记运用于低脂绵羊新品种的培育，同时该SNP所属AR也可作为一个重要候选功能基因应用于绵羊尾（臀）脂沉积分子机制的相关研究。

关键词: 脂尾性状 , 阿勒泰羊 , 多态性 , 基因型 , 标记基因

Abstract: 【Objective】 The objective of this paper is to study the polymorphism of the SNP located at the position of 60149247 on X chromosome between fat tail (rump) and thin tail breeds, and to provide the basic data for analysis of the mechanism of tail fat deposition in sheep.【Method】 Firstly, PCR-SSCP genotyping method was used to analyze genotypes among different tail types of sheep breeds, and definite allelic genes were cleared by PCR-direct sequencing, and the chi-square test was employed to detect the balance of genotype distribution in different breeds. Secondly, the SNP was mapped on sheep androgen receptor (AR) and the whole CDS sequences of AR were obtained by comparative genome and silicon cloning methods. Taking cDNA of Altay tail fat as DNA template, RT-PCR method was used to clone sheep AR spinning six exons, and sequence homology and phylogenetic tree were analyzed by bioinformatics methods. 【Result】The results of PCR-SSCP showed that there is no polymorphism in fat-tail sheep breeds (Hu sheep and Altay sheep), and the genotypes of the two breeds are all GG genotype. And there are three genotypes (GG, GA and AA) in thin-tail sheep populations (Chinese Merino sheep and Suffolk sheep), and the frequency of GG genotype dropped to 10% and 21%, respectively. The whole CDS sequences of sheep AR were cloned by silicon cloning, and sheep AR containing the fragment from the 3rd exon to 8th exon of AR was successfully cloned in total RNA of Altay tail fat for the first time, and the sequences are completely the same to the sequences predicted by silicon cloning. The sequence alignment also showed that AR is highly conserved in mammalian animals, and the homology was up to 90% among different mammalian animals, and the phylogenetic tree analysis showed that sheep is relatively close to bovine, pigs and dolphin. 【Conclusion】This study was the first to identify a novel SNP in the 3rd intron of the sheep AR associated with the trait of fat tail, and it hinted that the SNP will be used as a practical application of molecular markers in low-fat sheep breeding. At the same time, AR will also serve as an important candidate gene in studying tail fat deposition in Sheep.

Key words: fat tail trait , Altay sheep , polymorphism , genotype , marker gene

甘尚权, 沈敏, 李欢, 梁耀伟, 杨井泉, 高磊, 刘守仁, 王新华. X染色体60149273位点在脂尾（臀）和瘦尾绵羊品种中的多态性及其基因定位[J]. 中国农业科学, 2013, 46(22): 4791-4799.

GAN Shang-Quan, SHEN Min, LI Huan, LIANG Yao-Wei, YANG Jing-Quan, GAO Lei, LIU Shou-Ren, WANG Xin-Hua. Polymorphism of the 60149273th Loci on X Chromosome Among Fat Tail and Thin Tail Breeds and Its Gene Mapping[J]. Scientia Agricultura Sinica, 2013, 46(22): 4791-4799.

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参考文献

[1]王乐, 胡昕, 薛正芬, 张敏, 叶尔兰, 赛迪古丽•赛买提, 巴合提•吾拉孜汗, 庞静, 郑文新. 新疆阿勒泰地区阿勒泰羊毛绒品质分析. 家畜生态学报, 2012, 33(5): 39-41.

Wang Y, Hu X, Xue Z F, Zhang M, Ye E L, Saidigulisaimaiti, Barhetiwulazihan, Pang J, Zheng W X. Quality investigation and analysis of Altay sheep wool and down hair in Xinjiang Altay Region. Acta Ecologiae Animalis Domastici, 2012, 33(5): 39-41. (in Chinese)

[2]李金保, 别克•木哈买提, 库拉西, 努尔古丽. 地方良种阿勒泰羊. 新疆畜牧业, 2008, 1: 31-33.

Li J B, Biekemuhamaiti, Kulaxi，Nuerguli. Local improved varieties Altay sheep. Xinjiang Husbandry, 2008, 1: 31-33. (in Chinese)

[3]Momani-Shaker M, Abdullah A Y, Kridli R T, Blaha J, Sada I, Sovjak R. Fattening performance and carcass value of Awassi ram lambs, F1 crossbreds of Romanov×Awassi and Charollais×Awassi in Jordan. Czech Journal of Animal Science, 2002, 47(10): 429-438.

[4] Khaldari M K, Afzalzadeh A, Salehi A. Growth and carcass characteristics of crossbred progeny from lean tailed and fat tailed sheep breeds. South African Journal of Animal Science, 2007, 37(1): 51-56.

[5]许莲萍, 王文均, 郭晓昭, 姚刚. 断奶对阿勒泰羔羊血液生理生化指标的影响. 新疆农业大学学报, 2007, 112(2): 71-73.

Xu L P, Wang W J, Guo X Z, Yao G. Effect of weaning on the blood physiological and biochemical indexes of Altay lambs. Journal of Xinjiang Agricultural University, 2007, 112(2): 71-73. (in Chinese)

[6]刘志强, 徐郁哉, 石玉珂, 贝念湘, 佟梅岭, 李静敏, 阿尔肯, 李英, 牙合牙. 阿勒泰羊生理生化常值测定. 新疆农业大学学报, 1987, 3: 7-11.

Liu Z Q, Xu Y Z, Shi Y K, Bei N X, Tong M L, Li J M, Aerken, Li Y, Yaheya. Determination of physiological and biochemical common values on Altay sheep. Journal of Xinjiang Agricultural University, 1987, 3: 7-11. (in Chinese)

[7]魏彬, 刘志强, 贝念湘, 李新平, 黄俊成, 叶林. 阿勒泰羊生理生化血液流变学常值. 草食家畜, 1997, 1: 41-44.

Wei B, Liu Z Q, Bei N X, Li X P, Huang J C, Ye L. Hemorheologic values of physiological and biochemical in Aletai sheep. Grass Feeding Livestock, 1997, 1: 41-44. (in Chinese)

[8]Moradi M H, Nejati-Javaremi A, Moradi-Shahrbabak M, Dodds K G, McEwan J C. Genomic scan of selective sweeps in thin and fat tail sheep breeds for identifying of candidate regions associated with fat deposition. http://www.biomed central.com/1471-2156/13/10.

[9]Hu Z L, Park C A, Wu X L，Reecy J M. Animal QTLdb: an improved database tool for livestock animal QTL/association data dissemination in the post-genome era. Nucleic Acids Research, 2013, 41 (1): D871-D879.

[10]Rohrer G A, Keele J W. Identification of quantitative trait loci affecting carcass composition in swine: I. Fat deposition traits. Joural of Animal Science, 1998, 76(9): 2247-2254.

[11]Rohrer G A, Keele J W. Identification of quantitative trait loci affecting carcass composition in swine:II. Muscling and wholesale product yield traits. Joural of Animal Science, 1998, 76(9): 2255-2262.

[12]Lu N Z, Wardell S E, Burnstein K L, Defranco D, Fuller P J, Giguere V, Hochberg R B, McKay L, Renoir J M, Weigel N L, Wilson E M, McDonnell D P, Cidlowski J A. International Union of Pharmacology. LXV. The pharmacology and classification of the nuclear receptor superfamily: glucocorticoid, mineralocorticoid, progesterone, and androgen receptors. Pharmacological Reviews, 58(4): 782-797.

[13]Roy A K, Lavrovsky Y, Song C S, Chen S, Jung M H, Velu N K, Bi B Y, Chatterjee B. Regulation of androgen action. Vitamins and Hormones, 1999, 55: 309-352.

[14]Mooradian A D, Morley J E, Korenman S G. Biological actions of androgens. Endocrine Reviews, 1987, 8 (1): 1-28.

[15]Heinlein C A, Chang C. The roles of androgen receptors and androgen-binding proteins in nongenomic androgen actions. Molecular Endocrinology, 2002, 16(10): 2181-2187.

[16]McInnes K J, Smith L B, Hunger N I, Saunders P T, Andrew R, Walker B R. Deletion of the androgen receptor in adipose tissue in male mice elevates retinol binding protein 4 and reveals independent effects on visceral fat mass and on glucose homeostasis. Diabetes, 2012, 61(5): 1072-1081.

[17]Graham T E，Yang Q，Bluher M，Hammarstedt A, Ciaraldi T P, Henry R R, Wason C J, Oberbach A, Jansson P A, Smith U, Kahn B B. Retinol binding protein 4 and insulin resistance in lean,obese,and diabetic subjects. The New England Journal of Medicine, 2006, 354(24): 2552-2563.

[18]Zajac J, Rana K, Fam B, Andrikopoulos S, MacLean H. Increased fat mass in androgen receptor knockout mice is caused by decreased physical activity with no change in food consumption//11th European Congress Endocrinology. European Society of Endocrinology, 2009: 668.

[19]Lessard J, Tchernof A. Interaction of the glucocorticoid and androgen receptors in adipogenesis. Chemistry and Biology, 2012, 19(9): 1079-1080.

[20]Zitzmann M, Gromoll J, von Eckardstein A, Nieschlag E. The CAG repeat polymorphism in the androgen receptor gene modulates body fat mass and serum concentrations of leptin and insulin in men. Diabetologia, 2003, 46(1): 31-39.

[21]毛达勇, 赵娟, 张吉才. 雄激素受体基因(CAG)n多态性与男性2型糖尿病血管并发症相关性. 分子诊断与治疗杂志, 2012, 4(4): 245-248.

Mao D Y, Zhao J, Zhang J C. Correlation between androgen receptor gene(CAG)n polymorphism and male type 2 diabetic vascular complications. Journal of Molecular Diagnostics and Therapy, 2012, 4(4): 245-248. (in Chinese)

[22]Matias P M, Carrondo M A, Coelho R, Thomaz M, Zhao X Y, Wegg A, Crusius K, Egner U, Donner P. Structural basis for the glucocorticoid response in a mutant human androgen receptor (AR(ccr)) derived from an androgen-independent prostate cancer. Journal of Medicinal Chemistry, 2002, 45(7): 1439-1446.

[23]Tilley W D, Buchanan G, Hickey T E, Bentel J M. Mutations in the androgen receptor gene are associated with progression of human prostate cancer to androgen independence. Clinical Cancer Research, 1996, 2(2): 277-285.

[24]Ellis J A, Stebbing M, Harrap S B. Polymorphism of the androgen receptor gene is associated with male pattern baldness. Journal of Investigative Dermatology, 2001, 116(3): 452-455.

[25]Sutherland R W, Wiener J S, Hicks J P, Marcelli M, Gonzales E T, Roth D R, Lamb D J. Androgen receptor gene mutations are rarely associated with isolated penile hypospadias. Journal of Urology, 1996, 156(Pt. 2): 828-831.

[26]Lobaccaro J M, Lumbroso S, Berta P, Chaussain J L, Sultan C. Complete androgen insensitivity syndrome associated with a de novo mutation of the androgen receptor gene detected by single strand conformation polymorphism. Journal of Steroid Biochemistry and Molecular Biology, 1993, 44(3): 211-216.

[27]Ferlin A, Bartoloni L, Rizzo G, Roverato A, Garolla A, Foresta C. Androgen receptor gene CAG and GGC repeat lengths in idiopathic male infertility. Molecular Human Reproduction, 2004, 10(6): 417-421.

[28]Castro-Nallar E, Bacallao K, Parada-Bustamante A, Lardone M C, López P V, Madariaga M, Valdevenito R, Piottante A, Ebensperger M, Castro A. Androgen receptor gene CAG and GGN repeat polymorphisms in Chileanmen with primary severe spermatogenic failure. Journal of Andrology, 2010, 31(6): 552-559.

[29]Tong D, Deng J, Sun H, Chen L, Wu X. The relationship between CAG repeat length polymorphism and infertility in Southern Chinese Han women. Journal of Endocrinological Investigation, 2010, 33(8): 559-563.

[30]Shah N A, Antoine H J, Pall M, Taylor K D, Goodarzi M O. Association of androgen receptor CAG repeat polymorphism and polycystic ovary syndrome. Journal Clinical Endocrinology Metabolism, 2008, 93(5): 1939-1945.

[31]Liu G L, Jiang S W, Xiong Y Z, Zheng R, Qu Y C. Association of PCR-RFLP polymorphisms of IGF2 gene with fat deposit related traits in pig resource family. Yi Chuan Xue Bao, 2003, 30(12): 1107-1112.

[32]Knoll A, Putnová L, Dvorák J, Cepica S. A NciI PCR-RFLP within intron 2 of the porcine insulin like growth factor 2(IGF2) gene. Animal Genetics, 2000, 31(2): 150-151.

[33]van Meurs J B, Schuit S C, Weel A E, van der Klift M, Bergink A P, Arp P P, Colin E M, Fang Y, Hofman A, van Duijn C M, van Leeuwen J P, Pols H A, Uitterlinden A G. Association of 5′ estrogen receptor alpha gene polymorphisms with bone mineral density, vertebral bone area and fracture risk. Human Molecular Genetics, 2003, 12(14): 1745-1754.

[34]Okura T, Koda M, Ando F, Niino N, Ohta S, Shimokata H. Association of polymorphisms in the estrogen receptor alpha gene with body fat distribution. International Journal of Obesity Related Metablisim Disords, 2003, 27(9): 1020-1027

[35]Chang K C. Critical regulatory domains in intron 2 of a porcine sarcomeric myosin heavy chain gene. Journal of Muscle Researchand Cell Motility, 2000, 21(5): 451-461.

[36] Hormuzdi S G, Penttinen R, Jaenisch R, Bornstein P. A gene-targeting approach identifies a function for the first intron in expression of the alpha1(I) collagen gene. Molecular Celluar Biology, 1998, 18(6): 3368-3375.