湖羊CNTF基因组织表达、SNPs扫描及其与生长性状关联分析  

Tissue Expression,SNPs Scanning of CNTF Gene and Its Association with Growth Traits in Hu Sheep(Ovis aries)

作　　者：程江博 张德印 张煜坤 宋其志 李晓龙 赵源 徐丹 张小雪[1] 李冲[1] 袁律峰 李发弟[2,3] 王维民 CHENG Jiang-Bo;ZHANG De-Yin;ZHANG Yu-Kun;SONG Qi-Zhi;LI Xiao-Long;ZHAO Yuan;XU Dan;ZHANG Xiao-Xue;LI Chong;YUAN Lyu-Feng;LI Fa-Di;WANG Wei-Min(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;College of Grassland Agricultural Science and Technology,Lanzhou University,Lanzhou 730000,China;Gansu Mutton Sheep Breeding Biotechnology Engineering Laboratory,Minqin 733300,China)

机构地区：[1]甘肃农业大学动物科学技术学院,兰州730070 [2]兰州大学草地农业科技学院,兰州730000 [3]甘肃省肉羊生物技术繁育工程实验室,民勤733300

出　　处：《农业生物技术学报》2022年第3期506-516,共11页Journal of Agricultural Biotechnology

基　　金：国家畜禽良种联合攻关(19200140);国家自然科学基金(31760651);中国科学院“西部之光”人才培养计划“西部青年学者”A类项目资助。

摘　　要：睫状神经营养因子(ciliary neurotrophic factor,CNTF)是影响肌肉发育的重要候选基因,可以编码具有多种生物学功能的细胞因子。本研究旨在探讨绵羊(Ovis aries)CNTF基因的组织表达规律,扫描其SNPs并在大规模群体中与生长性状进行关联分析。本研究以1399只系谱清晰、表型记录准确的湖羊(Ovis aries)群体为研究对象,利用qPCR技术检测了绵羊睫状神经营养因子基因在湖羊10个组织的表达量,采用混合池测序的方法扫描该基因SNPs,并与生长性状进行关联分析。研究结果发现,生长性状间存在较高的正相关,体重与体长、体高、胸围和管围等体尺指标相关性较高(P<0.01)。绵羊CNTF基因广谱表达,在心、肝、脾、肺、肾等10个组织中均有表达,其中在肝脏和瘤胃中表达量较高。在绵羊CNTF基因第1内含子中检测到了g.2576 C>G多态位点,表现为中度多态。性状关联分析结果显示,该多态位点与湖羊80~180 d阶段的体重、体高、体长、胸围、管围等生长性状呈显著关联(P<0.05)。其中,携带GG基因型个体的80、100、120、140、160 d体重和80、100、120 d体高以及160 d胸围显著高于携带CG基因型个体(P<0.05),携带GG基因型个体的140、160、180 d体高和80、100、140和180 d胸围以及各阶段管围均显著高于携带CG基因型和CC基因型个体(P<0.05)。本研究结果表明CNTF基因g.2576 C>G位点可作为候选分子标记用于湖羊生长性状的育种实践。Ciliary neurotrophic factor(CNTF)is a cytokine with multiple biological functions in vivo.It is an important candidate gene affecting muscle development.The aim of this study was to investigate the tissue expression pattern of CNTF gene in sheep(Ovis aries),scan its SNPs and analyze its association with growth traits in a large population.In this study,1399 Hu sheep with clear family tree and accurate phenotype records were raised in a specialized performance measurement house.Before morning feeding at 80,100,120,140,160 and 180 d,respectively,the weight of the sheep was weighed,and body length,body height,chest circumference and cannon circumference were measured at the same time.At the end of the assay period,whole blood samples of each sheep were collected by jugular vein for genomic DNA extraction.They were slaughtered immediately and tissue samples were taken.According to design the CNTF mRNA sequence specific primers,RNA from the collected tissue samples was extracted and reverse transcribed to cDNA,using qPCR technology to detect the CNTF gene expression in 10 tissues of Hu sheep.Genomic DNA was extracted from whole blood samples of the experimental population using genomic DNA extraction kit according to the instructions.Mixed DNA was used as the template to amplify the designed PCR primers.SNPs of the gene was scanned by mixed pool sequencing method,and the SPSS 23.0 software was used for correlation analysis with sheep growth traits.This results found a high positive association between growth traits,with a higher correlation between weight and body size indicators such as body length,body height,chest circumferences and cannon circumferences(P<0.01).The ovine CNTF gene was ubiquitously expressed,with expression observed in 10 tissues including heart,liver,spleen,lung,and kidney,with higher expression in liver and rumen.The sheep CNTF gene fragment was amplified and sequenced,a g.2576 C>G polymorphism locus was detected in intron 1 of the sheep CNTF gene.The SNP sites were genotyped by KASPar technique,

关 键 词：湖羊 SNP 睫状神经营养因子基因(CNTF) 生产性状 

分 类 号：S813.3[农业科学—畜牧学]