绵羊KITLG基因的表达及其多态性与产羔性状关联分析  

Expression and Polymorphisms of KITLG Gene and Their Association with Litter Size in Sheep (Ovis aries)

作　　者：张军霞[1,4] 乐祥鹏[2] 王维民[1,3] 谢云龙[2] 王志兰[2] 张娜娜[2] 胡瑞雪[2] 孙武[2] 邓颖[2] 李发弟[1,2,3] 喇永富 

机构地区：[1]甘肃农业大学动物科技学院,兰州730070 [2]兰州大学草地农业科技学院,兰州730020 [3]甘肃省肉羊繁育生物技术工程实验室,民勤陇原中天生物工程有限公司,民勤733300 [4]青海大学农牧学院,西宁810016

出　　处：《农业生物技术学报》2017年第6期893-900,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(No.31472072);国家肉羊现代肉羊产业技术体系建设专项(CARS-39)

摘　　要：繁殖性状作为绵羊重要的经济性状,其分子机理的研究极为重要。绵羊(Ovis aries)繁殖性能受到下丘脑-垂体-性腺生殖轴的调控,同时也受到许多的生长因子的调节,KIT的配体KITLG(KIT LIGAND,KL或KitL)作为重要的生长调节因子在卵泡发育中起很重要的作用。为了解绵羊KITLG基因表达特征、多态性及其与产羔性状的关联性,本研究利用Real-time PCR分析了KITLG基因在湖羊下丘脑、垂体、卵巢、心脏、肌肉、肺、肾脏、肝脏、脾脏、十二指肠、脂肪和瘤胃等组织中的表达水平,同时采用限制性长度片段多态性(restriction fragment length polymorphism,RFLP)技术检测了KITLG基因在湖羊和小尾寒羊群体中的多态性及其与产羔性状的关联性。结果表明,绵羊KITLG基因在上述检测的12个组织中均有表达,其中在卵巢中的表达量较高。通过PCR-RFLP分析,在第2和第5内含子上检测到g.38473C>T和g.47468C>T 2个突变位点。在小尾寒羊和湖羊群体中检测到的g.38473C>T位点上均有CC(优势基因型)和CT两种基因型,其中C为优势等位基因;湖羊群体的基因杂合度(heterozygosity,He)为0.89,纯合度(homozygosity,Ho)为0.11,有效等位基因数(effective allele number,Ne)和多态信息含量(polymorphic information content,PIC)分别为1.31和0.11;在小尾寒羊中分别为0.80、0.20、1.10和0.16,并且在该突变位点,两个群体均处于Hardy-Weinberg平衡状态。在g.47468C>T突变位点上检测到CT和TT(优势基因型)两种基因型,其中优势等位基因为T;在此位点湖羊群体中He、Ho、Ne及PIC分别为0.95、0.05、1.06、0.05;小尾寒羊群体中分别为0.91、0.09、1.10和0.09。两个群体在该位点均处于Hardy-Weinberg平衡状态。多态位点与绵羊产羔数的关联性分析表明,KITLG基因g.47468C>T位点与小尾寒羊产羔数显著相关(P<0.05)。本研究结果揭示了KITLG基因在绵羊卵巢中的表达量较高,且g.47468C>T位点与绵羊产羔数显著Reproductive performance is an important economic trait in sheep （Ovis aries）. The study of its molecular mechanism is also extremely important. Reproductive performance of sheep is regulated by the hypothalamic- pituitary- gonad reproductive axis, and is also controlled by many growth factors. KITLG acting as ligand of KIT （KIT LIGAND, KL or KitL）, as an important growth regulation factor, plays a key role in follicular development. The current study aimed to analyze the expression ofKJTLG gene and SNPs as well as their association with litter size in sheep, and to offer a reference data for the candidate gene research of reproduction trait in sheep. Therefore, Real time-PCR was conducted to investigate the expression patterns of KITLG gene in hypothalamus, pituitary, ovary, heart, muscle, lung, kidney, liver, spleen, duodenum, fat and rumen tissues of Hu sheep, and restriction fragment length polymorphism （RFLP） technologies were carried out to screen SNPs and analyze their association with litter size in sheep population of Hu sheep and Small tail Han sheep. The results revealed that KITLG gene expressed in heart, liver, spleen, lung, kidney, rumen, duodenum, muscle, fat, hypothalamus, pituitary, and the expression in ovary was high. According to PCR- RFLP technologies, two polymorphisms g.38473C〉T and g.47468C〉T2 were identified in intron 2 and 5 ofKITLG gene respectively. In g.38473C〉T locus, two genotypes, CC and CT were found in both Hu sheep and Small tail Han sheep. Of them, CC was the dominant genotype, and C was the dominant allele. In addition, the homozygosity （110）, heterozygosity （He）, effective allele number （Ne）, polymorphic information content （PIC） were 0.89, 0.11, 1.31 and 0.11 for Hu sheep, and were 0.80, 0.20, 1.10 and 0.16 for Small tail Han sheep, respectively. The X2 test indicated that Small tail Han sheep and Hu sheep populations were in Hardy-Weinberg equilibrium. Same as g.38473C〉T locus, two genotypes, CT and TT were identified in g.4746gC〉T loc

关 键 词：KITLG基因 单核苷酸多态性 产羔数 基因表达 绵羊 

分 类 号：S813.3[农业科学—畜牧学]