Leptin基因多态性与绵羊季节性发情的相关分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (9): 189-194.

Leptin基因多态性与绵羊季节性发情的相关分析

李辉^1,2,3,卢守亮^1,2,3,万鹏程^1,3,代蓉^1,3,李良远^1,2,3,孟浩浩^1,2,3,石国庆^1,3

1. 新疆农垦科学院畜牧兽医研究所，石河子 832000;
2. 石河子大学动物科技学院，石河子 832003;
3. 新疆兵团绵羊繁育生物技术重点实验室，石河子 832000

收稿日期:2014-02-02 出版日期:2014-09-15 发布日期:2014-09-07
作者简介:李辉，男，硕士研究生，研究方向：动物遗传育种与繁殖；E-mail：huili223344@163.com
基金资助:
国家科技支撑计划（2011BAD28B05-1-1），新疆兵团育种攻关（2011BA006），新疆兵团绵羊繁育生物技术重点实验室开放课题（2013KLS07）

Analysis of the Relationship Between Polymorphism of Leptin and Seasonal Estrous in Sheep

Li Hui^1,2,3,Lu Shouliang^1,2,3,Wan Pengcheng^1,3,Dai Rong^1,3,Li Liangyuan^1,2,3,Meng Haohao^1,2,3,Shi Guoqing^1,3

1. Institute of Animal Husbandry and Veterinary，Xinjiang Academy of Agricultural and Reclamation Sciences，Shihezi University，Shihezi 832000;
2. College of Animal Science and Technology，Shihezi 832003;
3. Key Lab of Sheep Breeding and Reproduction，Xinjiang Agroreclamation Academy of Sciences，Shihezi 832000

Received:2014-02-02 Published:2014-09-15 Online:2014-09-07

摘要/Abstract

摘要： 根据leptin基因在GenBank中的已知序列设计两对引物，采用PCR-SSCP技术在常年发情的湖羊和季节性发情的阿勒泰羊群体中进行单核苷酸多态性（SNPs）检测，对筛查到的SNP位点进行基因型与绵羊季节性发情的关联分析。结果表明，与湖羊相比，阿勒泰羊leptin基因第1内含子上有3个连续碱基TTG的插入和C/T碱基突变；第3外显子3上发生G/T碱基突变，编码氨基酸由缬氨酸变成亮氨酸。Leptin外显子2扩增片段上检测到AA、AB、BB三种基因型，BB基因型在阿勒泰羊群体中属于优势基因型；对两个群体进行基因型频率独立性χ2检验，差异极显著（P<0.001），说明BB基因型是影响季节性发情的有利基因型。研究结果提示，绵羊品种中Leptin基因序列的差异性可能是造成绵羊季节性发情的原因之一，可作为常年发情绵羊品种选育的辅助标记。

关键词: 绵羊, Leptin, 季节性发情, PCR-SSCP

Abstract: According to the known sequence of leptin gene from GenBank, two primers were designed. The single nucleotide polymorphisms（SNPs）of the nonseasonal estrus Hu sheep and seasonal estrus Altay sheep were detected using PCR-SSCP assay. Then, the screening SNPs was subjected to the relative analysis of genotype and seasonal estrus. The results showed that three consecutive bases TTG inserted and C / T mutated intron 1 of leptin in Altay sheep, and G / T mutation in exon 3 resulted in the replacement of valine by leucine in contrast with the Hu sheep. Three genotypes AA, AB, BB were detected on the amplified fragment of the exon 2, the genotype BB was dominant in Altay. The genotype distributions were significantly different（P<0.001）between two groups, especially demonstrated that genotype BB was benefit for seasonal estrus. In conclusion, the sequence difference of Leptin in different sheep breeds may be one possible reason leading to nonseasonal or seasonal estrus, which can be used as an assistant marker for nonseasonal estrus breeding.

Key words: Sheep , Leptin , Seasonal estrous , PCR-SSCP

李辉,卢守亮,万鹏程,代蓉,李良远,孟浩浩,石国庆. Leptin基因多态性与绵羊季节性发情的相关分析[J]. 生物技术通报, 2014, 0(9): 189-194.

Li Hui,Lu Shouliang,Wan Pengcheng,Dai Rong,Li Liangyuan,Meng Haohao,Shi Guoqing. Analysis of the Relationship Between Polymorphism of Leptin and Seasonal Estrous in Sheep[J]. Biotechnology Bulletin, 2014, 0(9): 189-194.

参考文献

［1］Zhang Y, Proenca R, Maffei M, et al. Positional cloning of the mouse obese gene and its human homologue［J］. Nature, 1994, 372（6505）：425-432.
［2］Perucatti A, Di Meo GP, Vallinoto M, et al. FISH-mapping of LEP and SLC26A2 genes in sheep, goat and cattle R-banded chromoso-mes：comparison between bovine, ovine and caprine chromosome 4（BTA4/OAR4/CHI4）and human chromosome 7（HSA7）［J］. Cytogenetic and Genome Research, 2006, 115（1）：7-9.
［3］Rahmouni K, Haynes WG. Leptin and the central neural mechanisms of obesity hypertension［J］. Drugs Today, 2002, 38（12）：807-817.
［4］李艳利, 彭焕文, 杨雁, 等. leptin研究进展［J］.安徽农业科学, 2012, 40（11）：6363-6367.
［5］Kale DS, Yadav BR, Mukherjee A, et al. Exploring DNA polymorp-hisms of Leptin gene within Indian water buffaloes［J］. Journal of Advanced Veterinary Research, 2013, 3（1）：20-26.
［6］Wabitsch M, Jensen PB, Blum WF, et al. Insulin and cortisol promote leptin production in cultured human fat cells［J］. Diabetes, 1996, 45（10）：1435-1438.
［7］Tanpure T, Dubey PK, Singh KP, et al. PCR-SSCP analysis of leptin gene and its association with milk production traits in river buffalo（Bubalus bubalis）［J］. Tropical Animal Health and Production, 2012, 44（7）：1587-1592.
［8］Jin L, Zhang S, Burguera B G, et al. Leptin and leptin receptor expression in rat and mouse pituitary cells［J］. Endocrinology, 2000, 141（1）：333-339.
［9］Chagnon YC, Wilmore JH, Borecki IB, et al. Associations between the leptin receptor gene and adiposity in middle-aged Caucasian males from the HERITAGE family study［J］. Journal of Clinical Endocrinology & Metabolism, 2000, 85（1）：29-34.
［10］杨静, 杨琳. 瘦素对繁殖的影响［J］. 广东饲料, 2013, 22（7）：25-26.
［11］Datta S, Adikari A, Chauhan A, et al. Nucleotide sequence variation in leptin gene of Murrah buffalo（Bubalus bubalis）［J］. Exploratory Animal and Medical Research, 2012, 2（2）：130-136.
［12］Sullivan SD, Howard LC, Clayton AH, et al. Serotonergic activation rescues reproductive function in fasted mice；Does serotonin mediate the metabolic effects of leptin on reproduction?［J］. Biology of Reproduction, 2002, 66（6）：1702-1706.
［13］Schneider JE, Blum RM, Wade GN. Metabolic control of food intake and estrous cycles in Syrian hamsters. I. Plasma insulin and leptin［J］. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology, 2000, 278（2）：476-485.
［14］Sosa C, Carriquiry M, Chalar C, et al. Endometrial expression of leptin receptor and members of the growth hormone—Insulin-like growth factor system throughout the estrous cycle in heifers［J］. Animal Reproduction Science, 2010, 122（3）：208-214.
［15］杨海兵, 杨胜林, 王昌贵, 等. 贵州黑白花奶牛 Leptin 基因启动子区多态性与产奶性状关联分析［J］. 黑龙江畜牧兽医, 2013（2）：46-49.
［16］Dubey PP, Sharma A, Gour DS, et al. Leptin gene polymorphism in Indian Sahiwal cattle by single strand conformation polymorphism（SSCP）［J］. South African Journal of Animal Science, 2008, 38（2）：131-135.
［17］黄丹丽, 陈仁余, 杨章平, 等. 4个绵羊品种Leptin基因部分片段的SNPs多态性与生长发育性状的相关分析［J］. 畜牧兽医学报, 2008（12）：1640-1646.
［18］Barzehkar R, Salehi A, Mahjoubi F. Polymorphisms of the ovine leptin gene and its association with growth and carcass traits in three Iranian sheep breeds［J］. Iranian Journal of Biotechnology（IJB）, 2009, 7（4）：241-246.
［19］Hormuzdi SG, Penttinen R, Jaenisch R, et al. A gene-targeting approach identifies a function for the first intron in expression of the α1（I）collagen gene［J］. Molecular and Cellular Biology, 1998, 18（6）：3368-3375.
［20］Chang KC. Critical regulatory domains in intron 2 of a porcine sarcomeric myosin heavy chain gene［J］. Journal of Muscle Research & Cell Motility, 2000, 21（5）：451-461.
［21］Bahrami A, Behzadi S, Miraei-Ashtiani SR, et al. Genetic polymorphisms and protein structures in growth hormone, growth hormone receptor, ghrelin, insulin-like growth factor 1 and leptin in Mehraban sheep［J］. Gene, 2013, 527（1）：397-404.
［22］刘宝凤, 周沙沙, 王景霖, 等. LEP 和LEPR基因在不同脂尾型绵羊脂肪组织中的mRNA表达研究［J］. 畜牧兽医学报, 2013, 44（7）：1014-1022.
［23］邹果, 张亚平, 于黎. 瘦蛋白（leptin）自然选择和适应性进化研究概述［J］. 科学通报, 2013, 58（16）：1473-1482.
［24］Yang J, Wang ZL, Zhao XQ, et al. Natural selection and adaptive evolution of leptin in the ochotona family driven by the cold environmental stress［J］. PLoS One, 2008, 3（1）：e1472.

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