贵州半细毛羊促生长激素分泌素受体基因SNPs筛查与变异分析

中国畜牧兽医

贵州半细毛羊促生长激素分泌素受体基因SNPs筛查与变异分析

张琼娣¹，宋德荣¹，周大荣¹，彭华¹，刘若余²，吴蕊汝¹，江兴美¹，吴萍¹

（1.毕节市畜牧兽医科学研究所，贵州毕节 551700；2.贵州大学动物科学学院，贵州花溪 550025）

收稿日期:2013-09-24 出版日期:2014-04-20 发布日期:2014-05-27
通讯作者: 宋德荣（1967—），男，学士，研究员，研究方向：动物遗传育种。E-mail：sdr0857@126.com
作者简介:张琼娣（1970—），女，贵州人，硕士，助理研究员，研究方向：分子生物学与繁殖。
基金资助:
国家现代农业产业技术体系专项资金（CARS-40-30、CARS-40-46）；贵州省农业动植物育种专项（黔农育专字［2011］016号）；毕节地区科技计划项目（毕科合字［2010］05号）；贵州省科技厅、毕节地区行署、中科院昆明分院科技合作项目（省地院合2010-05）；毕节试验区肉羊产业发展研究人才团队（毕人领办通［2013］7号）。

Screening the GHSR Gene for SNPs and Analyzing the Mutations in Guizhou Semi-fine Wool Sheep

ZHANG Qiong-di¹, SONG De-rong¹，ZHOU Da-rong¹，PENG Hua¹，LIU Ruo-yu²，WU Rui-ru¹，JIANG Xing-mei¹，WU Ping¹

（1.Animal Husbandry and Veterinary Science Institute of Bijie City, Bijie 551700，China；2.College of Animal Science, Guizhou University, Huaxi 550025，China）

Received:2013-09-24 Online:2014-04-20 Published:2014-05-27

摘要/Abstract

摘要： 通过研究促生长激素分泌素受体（growth hormone secretagogue receptor，GHSR）基因多态性，为贵州半细毛羊选种选育提供进一步的分子生物学依据。试验选取36只贵州半细毛羊构建DNA池，设计1对引物扩增其GHSR基因第2外显子及第1、2内含子部分序列并进行双向测序，对SNPs位点等位基因频率进行估算，利用在线生物信息学软件分析SNPs位点对GHSR基因RNA二级结构的影响。结果发现，在贵州半细毛羊GHSR基因中筛选到2个SNPs：T70G和G229A，均为同义突变，SNPs位点导致GHSR基因mRNA二级结构发生变化。GHSR基因多态性可能影响贵州半细毛羊的生长。

关键词: 贵州半细毛羊; 促生长激素分泌素受体基因; SNP; DNA池

Abstract: By studying the GHSR gene polymorphism, A basis of selection and breeding was provide on molecular biology of Guizhou Semi-fine Wool sheep. Guizhou Semi-fine Wool sheep were used to construct DNA pools in this study, the complete sequences of exon 2 and part of intron 1 and intron 2 region in GHSR gene were obtained by designing one pair of specific primers. The amplified PCR products would be directly purified and conducted bidirectional sequences subsequently. DNAStar and BLAST software were used to identify polymorphisms of GHSR gene. Moreover, the impact of SNPs on secondary structure of RNA, secondary structure of GHSR protein would be analyzed by the bioinformatics software. The results showed that two SNPs (T70G and G229A) were found in GHSR gene which were two synonymous mutations. The SNPs led to the changes of RNA secondary structure. GHSR gene polymorphism might affect the growth of Guizhou Semi-fine Wool sheep.

Key words: Guizhou Semi-fine Wool sheep; GHSR gene; SNP; DNA pools

张琼娣，宋德荣，周大荣，彭华，刘若余，吴蕊汝，江兴美，吴萍. 贵州半细毛羊促生长激素分泌素受体基因SNPs筛查与变异分析[J]. 中国畜牧兽医.

ZHANG Qiong-di, SONG De-rong，ZHOU Da-rong，PENG Hua，LIU Ruo-yu，WU Rui-ru，JIANG Xing-mei，WU Ping. Screening the GHSR Gene for SNPs and Analyzing the Mutations in Guizhou Semi-fine Wool Sheep[J]. .

参考文献

1 方梅霞，李莹，徐海平，等. GHRL及其受体GHSR基因多态性与鸭生长及屠体性状的关联性［J］. 畜牧兽医学报，2011,42(1):18～24.
2 李丽娟,申小云.贵州半细毛羊的培育历程与养殖现状［J］.贵州农业科学, 2010,38(11):182～184.
3 李敬瑞,丁远华,向程举，等.猪DAZ基因的DNA池测序分析［J］.中国畜牧兽医, 2011, 38(8): 60～62.
4 朱文奇,李慧芳,宋卫涛，等.生长素基因 Ghrelin对高邮鸭产蛋性能的遗传效应［J］.四川农业大学学报，2009,27(3):354～359.
5 杨永强，焦仁刚,龚俞,等.DNA池快速筛查牛SNPs STAM基因及估算等位基因频率［J］.生物技术,2012,22(5)：57～60.
6 陈荣,雷莉萍.贵州半细毛羊几个性状的观察分析［J］.贵州农业科学,1992, 1:34～35.
7 张宝,陈宏,赵苗，等.GHSR 基因多态性与南阳牛生长关系研究［A］.细胞·生命·健康——第十一届中国细胞生物学学术大会暨2009西安细胞生物学国际会议论文集［C］. 2009.
8 高风英，王欢，叶星，等. 荷那龙罗非鱼两种GHSR基因的克隆与序列分析［J］. 广东海洋大学学报，2011,31(4):7～11.
9 黄希贵,焦保卫,王德寿.生长激素促分泌素受体(GHSR)的发现及影响［J］. 动物学杂志, 2004, 39(2): 84～88.
10 谢海强,杨永强,孙岩岩，等.贵州白山羊和黔北麻羊TFAM基因部分外显子多态性研究［J］. 广东农业科学, 2013, 40(16): 138～140.
11 Gnanapavan S, Kola B, Bustin S A, et al. The tissue distribution of the mRNA of ghrelin and subtypes of its receptor, GHSR, in humans［J］. Journal of Clinical Endocrinology & Metabolism, 2002, 87(6): 2988.
12 Hayashida T, Murakami K, Mogi K, et al. Ghrelin in domestic animals: Distribution in stomach and its possible role［J］. Domestic Animal Endocrinology, 2001, 21(1): 17～24.
13 Kojima M, Hosoda H, Date Y, et al. Ghrelin is a growth-hormone-releasing acylated peptide from stomach［J］. Nature, 1999, 402(6762): 656～660.
14 Kojima M, Kangawa K. Ghrelin, an orexigenic signaling molecule from the gastrointestinal tract［J］. Current Opinion in Pharmacology, 2002, 2(6): 665～668.
15 Miyasaka K, Hosoya H, Sekime A, et al. Association of ghrelin receptor gene polymorphism with bulimia nervosa in a Japanese population［J］. Journal of Neural Transmission, 2006, 113(9): 1279～1285.
16 Nakazato M, Murakami N, Date Y, et al. A role for ghrelin in the central regulation of feeding［J］. Nature, 2001, 409(6817): 194～198.
17 Shuto Y, Shibasaki T, Otagiri A, et al. Hypothalamic growth hormone secretagogue receptor regulates growth hormone secretion, feeding, and adiposity［J］. Journal of Clinical Investigation, 2002, 109(11): 1429～1436.
18 Tschop M, Smiley D L, Heiman M L. Ghrelin induces adiposity in rodents［J］. Nature, 2000, 407(6806): 908～913.
19 Wang H J, Geller F, Dempfle A, et al. Ghrelin receptor gene: Identification of several sequence variants in extremely obese children and adolescents, healthy normal-weight and underweight students, and children with short normal stature［J］. Journal of Clinical Endocrinology & Metabolism, 2004, 89(1): 157～162.
20 Wren A M, Small C J, Abbott C R，et al. Ghrelin causes hyperphagia and obesity in rats［J］. Diabetes, 2001, 50(11): 2540～2547.
21 Zigman J M, Nakano Y, Coppari R, et al. Mice lacking ghrelin receptors resist the development of diet-induced obesity［J］. Journal of Clinical Investigation, 2005, 115(12): 3564～3572.