To establish a rapid, accurate and economical real-time PCR assay system based on TaqMan probe technology for the detection of genetic variations of single nucleotide polymorphisms （SNPs） in myestatin （MSTN） gene,...To establish a rapid, accurate and economical real-time PCR assay system based on TaqMan probe technology for the detection of genetic variations of single nucleotide polymorphisms （SNPs） in myestatin （MSTN） gene, a pair of TaqMan probes were designed on the polymorphism loci of MSTN gene and used in PCR reaction system for SNP genotyping. Meanwhile, an association study was performed between MSTN genotypes and growth traits of Tan sheep, including birth weight, weaning weight, 3-month weight, and 6-month weight. The results showed that rs417816017 locus of MSTN gene in Tan sheep had two genotypes ： YY and XY. The individuals with genotype XY had a growth advantage over the ones with genotype YY. The results indicate that TaqMan probe-based real-time PCR assay can be used to detect the genotype of MSTN gene, which will provide candidate genes for breeding of Tan sheep.展开更多
As a member of the Frizzled family, Frizzled3 (FZD3) is a receptor of the canonical Wnt signaling pathway and plays a vital role in mammalian hair follicle developmental processes. However, its effects on wool traits ...As a member of the Frizzled family, Frizzled3 (FZD3) is a receptor of the canonical Wnt signaling pathway and plays a vital role in mammalian hair follicle developmental processes. However, its effects on wool traits are not clear. The objectives of this study were to identify the single nucleotide polymorphisms (SNPs) and the expression patterns of FZD3 gene, and then to determine whether it affected wool traits of Chinese Merino sheep (Xinjiang Type) or not. PCR-single stranded conformational polymorphism (PCR-SSCP) and sequencing were used to identify mutation loci, and general linear model (GLM) with SAS 9.1 was used for the association analysis between wool traits and SNPs. Quantitative real-time PCR (qRT-PCR) was used to investigate FZD3 gene expression levels. The results showed that six exons of FZD3 gene were amplified and two mutation loci were identified in exon 1 (NC_019459.2: g.101771685 T>C (SNP1)) and exon 3 (NC_019459.2: g.101810848, A>C (SNP2)), respectively. Association analysis showed that SNP1 was significantly associated with mean fiber diameter (MFD)(P=0.04) and live weight (LW)(P=0.0004), SNP2 was significantly associated with greasy fleece weight (GFW)(P=0.04). The expression level of FZD3 gene in skin tissues of the superfine wool (SF) group was significantly lower (P<0.05) than that of the fine wool (F) group. Moreover, it had a higher expression level (P<0.01) in skin tissues than in other tissues of Chinese Merino ewes. While, its expression level had a fluctuant expression in skin tissues at different developmental stages of embryos and born lambs, with the highest expression levels (P<0.01) at the 65th day of embryos. Our study revealed the genetic relationship between FZD3 variants and wool traits and two identified SNPs might serve as potential and valuable genetic markers for sheep breeding and lay a molecular genetic foundation for sheep marker-assisted selection (MAS).展开更多
Background： In farm animals, mitochondrial DNA（mtDNA） effect on economic performance remains hot-topic for breeding and genetic selection. Here, 53 maternal lineages of Small-tailed Han sheep were used to investiga...Background： In farm animals, mitochondrial DNA（mtDNA） effect on economic performance remains hot-topic for breeding and genetic selection. Here, 53 maternal lineages of Small-tailed Han sheep were used to investigate the association of mitochondrial DNA variations and the lambing litter size.Results： Sequence sweeping of the mitochondrial coding regions discovered 31 non-synonymous mutations, and the association study revealed that T7719G in mt DNA t RNA-Lys gene was associated with litter size（P 〈 0.05）,manifesting 0.29 lambs per litter between the G and T carriers. Furthermore, using the mixed linear model, we assayed the potential association of the ovine litter size and haplogroups and multiple-level mtDNA haplotypes,including general haplotypes, assembled haplotypes of electron transport chain contained sequences（H-ETC）,mitochondrial respiratory complex contained sequences（H-MRC） and mitochondrial genes（H-gene, including polypeptide-coding genes, rRNA genes and tRNA genes）. The strategy for assembled mitochondrial haplotypes was proposed for the first time in mtDNA association analyses on economic traits, although none of the significant relations could be concluded（P 〉 0.05）. In addition, the nuclear major gene BMPR1B was significantly correlated with litter size in the flock（P 〈 0.05）, however, did not interact with mtDNA T7719G mutation（P 〉 0.05）.Conclusions： Our results highlight mutations of ovine mitochondrial coding genes, suggesting T7719G in tRNA-Lys gene be a potentially useful marker for selection of sheep litter size.展开更多
旨在利用全基因组关联分析探寻敖汉细毛羊羊毛性状新的分子标记和候选基因。本研究采集1~2周岁的健康敖汉细毛羊耳组织与羊毛作为试验素材,其中,母羊248只,公羊81只,总计329只。羊毛进行性状测定(包括纤维直径、自然长度、伸直长度、伸...旨在利用全基因组关联分析探寻敖汉细毛羊羊毛性状新的分子标记和候选基因。本研究采集1~2周岁的健康敖汉细毛羊耳组织与羊毛作为试验素材,其中,母羊248只,公羊81只,总计329只。羊毛进行性状测定(包括纤维直径、自然长度、伸直长度、伸直率),并对表型数据进行描述性统计和相关性分析。利用绵羊40K液相SNP芯片对全部个体进行基因分型。使用Plink 1.07软件对芯片数据进行质控,使用GCTA软件和PopLDdecay软件对质控数据进行群体结构分析。利用GMEMA混合线性模型对4种羊毛性状进行了全基因组关联分析(genomewide association study,GWAS),利用在线软件对候选基因进行GO和KEGG富集分析。质控后得到329只个体的30079个SNPs位点用于后续分析。通过GWAS分析筛选出4个在全基因组上显著相关的SNPs位点可能影响羊毛经济性状,分别位于1号、6号及8号染色体上。筛选出9个在染色体水平上显著相关的SNPs位点可能对羊毛性状具有潜在意义,分别位于3、5、8、11、18、21、22、25号染色体,寻找到39个可能影响羊毛性状的候选基因。本研究结果为后续探究敖汉细毛羊羊毛性状的遗传机制及分子育种标记开发提供重要参考。展开更多
为了研究阿勒泰大尾羊和小尾寒羊不同脂肪组织及其他组织之间基因表达的差异,探讨脂肪与肥胖相关基因(fat mass and obesity associated gene,FTO)与绵羊体脂沉积的相关性。随机各选择6只6月龄雄性阿勒泰大尾羊和小尾寒羊,禁食24h后屠宰...为了研究阿勒泰大尾羊和小尾寒羊不同脂肪组织及其他组织之间基因表达的差异,探讨脂肪与肥胖相关基因(fat mass and obesity associated gene,FTO)与绵羊体脂沉积的相关性。随机各选择6只6月龄雄性阿勒泰大尾羊和小尾寒羊,禁食24h后屠宰,分别采集下丘脑、海马、背最长肌、心肌和脂肪组织样本,应用冰冻组织切片技术,测定脂肪细胞的面积,用real-time PCR检测FTO基因在各组织中的表达水平。结果显示,阿勒泰大尾羊尾脂脂肪细胞面积极显著高于肾周脂脂肪细胞面积(P<0.01)。阿勒泰大尾羊与小尾寒羊下丘脑和海马FTO基因表达水平无显著差异,背最长肌、心肌FTO基因表达水平阿勒泰大尾羊的极显著高于小尾寒羊的(P<0.01),肾周脂中FTO基因表达水平阿勒泰大尾羊的显著高于小尾寒羊的(P<0.05)。阿勒泰大尾羊肾周脂、心周脂和尾脂中FTO基因表达水平在肾周脂、心周脂、尾脂之间无显著差异。结果表明,FTO基因表达水平在阿勒泰大尾羊和小尾寒羊之间存在品种间的差异。展开更多
本研究旨在利用全基因组关联分析方法(genome-wide association studies,GWAS)挖掘影响绵羊体重性状的遗传标记和功能基因。采用Illumina OvineSNP50BeadChip中密度商业化羊芯片,对3个品种(苏尼特羊、德国肉用美利奴羊和杜泊羊)共计329...本研究旨在利用全基因组关联分析方法(genome-wide association studies,GWAS)挖掘影响绵羊体重性状的遗传标记和功能基因。采用Illumina OvineSNP50BeadChip中密度商业化羊芯片,对3个品种(苏尼特羊、德国肉用美利奴羊和杜泊羊)共计329只绵羊的体重性状(初生重、断奶重、6月龄重、断奶前日增重、断奶后日增重、日增重)进行GWAS分析。统计分析和基因注释基于TASSEL软件、混合线性模型和2012年10月公布的最新版绵羊基因组Ovis_aries_v3.1序列信息。结果表明,有10个SNPs位点在全基因组显著水平上与断奶后日增重相关,部分位点分布在羊注释基因内(MEF2B、RFXANK等);除此之外还检测到22个SNPs在染色体显著水平上与其他体重性状存在相关性,获得一批重要的候选基因。这些基因对绵羊体重性状功能基因的挖掘具有重要的理论意义和参考价值。展开更多
基金Supported by Special Fund for Science and Technology Development of Ningxia Hui Autonomous Region(2012ZDN1001)Domestic Animal Germplasm Resources Sharing Platform Project(2005DKA21101)
文摘To establish a rapid, accurate and economical real-time PCR assay system based on TaqMan probe technology for the detection of genetic variations of single nucleotide polymorphisms （SNPs） in myestatin （MSTN） gene, a pair of TaqMan probes were designed on the polymorphism loci of MSTN gene and used in PCR reaction system for SNP genotyping. Meanwhile, an association study was performed between MSTN genotypes and growth traits of Tan sheep, including birth weight, weaning weight, 3-month weight, and 6-month weight. The results showed that rs417816017 locus of MSTN gene in Tan sheep had two genotypes ： YY and XY. The individuals with genotype XY had a growth advantage over the ones with genotype YY. The results indicate that TaqMan probe-based real-time PCR assay can be used to detect the genotype of MSTN gene, which will provide candidate genes for breeding of Tan sheep.
基金supported by the National Natural Science Foundation of China (31360543 and 31760655)the earmarked fund for the China Agriculture Research System (CARS-39)+1 种基金the Open Project of Key Laboratory of Genetics Breeding and Reproduction of Xinjiang Cashmere and Wool Sheep, China (2016D03017)the Postdoctoral Science Foundation, China (2017M623287)
文摘As a member of the Frizzled family, Frizzled3 (FZD3) is a receptor of the canonical Wnt signaling pathway and plays a vital role in mammalian hair follicle developmental processes. However, its effects on wool traits are not clear. The objectives of this study were to identify the single nucleotide polymorphisms (SNPs) and the expression patterns of FZD3 gene, and then to determine whether it affected wool traits of Chinese Merino sheep (Xinjiang Type) or not. PCR-single stranded conformational polymorphism (PCR-SSCP) and sequencing were used to identify mutation loci, and general linear model (GLM) with SAS 9.1 was used for the association analysis between wool traits and SNPs. Quantitative real-time PCR (qRT-PCR) was used to investigate FZD3 gene expression levels. The results showed that six exons of FZD3 gene were amplified and two mutation loci were identified in exon 1 (NC_019459.2: g.101771685 T>C (SNP1)) and exon 3 (NC_019459.2: g.101810848, A>C (SNP2)), respectively. Association analysis showed that SNP1 was significantly associated with mean fiber diameter (MFD)(P=0.04) and live weight (LW)(P=0.0004), SNP2 was significantly associated with greasy fleece weight (GFW)(P=0.04). The expression level of FZD3 gene in skin tissues of the superfine wool (SF) group was significantly lower (P<0.05) than that of the fine wool (F) group. Moreover, it had a higher expression level (P<0.01) in skin tissues than in other tissues of Chinese Merino ewes. While, its expression level had a fluctuant expression in skin tissues at different developmental stages of embryos and born lambs, with the highest expression levels (P<0.01) at the 65th day of embryos. Our study revealed the genetic relationship between FZD3 variants and wool traits and two identified SNPs might serve as potential and valuable genetic markers for sheep breeding and lay a molecular genetic foundation for sheep marker-assisted selection (MAS).
基金supported by the Project of Science and Technology of Hebei Province(15226308D)the National Key Basic Research Program of China(2014CB138500)
文摘Background： In farm animals, mitochondrial DNA（mtDNA） effect on economic performance remains hot-topic for breeding and genetic selection. Here, 53 maternal lineages of Small-tailed Han sheep were used to investigate the association of mitochondrial DNA variations and the lambing litter size.Results： Sequence sweeping of the mitochondrial coding regions discovered 31 non-synonymous mutations, and the association study revealed that T7719G in mt DNA t RNA-Lys gene was associated with litter size（P 〈 0.05）,manifesting 0.29 lambs per litter between the G and T carriers. Furthermore, using the mixed linear model, we assayed the potential association of the ovine litter size and haplogroups and multiple-level mtDNA haplotypes,including general haplotypes, assembled haplotypes of electron transport chain contained sequences（H-ETC）,mitochondrial respiratory complex contained sequences（H-MRC） and mitochondrial genes（H-gene, including polypeptide-coding genes, rRNA genes and tRNA genes）. The strategy for assembled mitochondrial haplotypes was proposed for the first time in mtDNA association analyses on economic traits, although none of the significant relations could be concluded（P 〉 0.05）. In addition, the nuclear major gene BMPR1B was significantly correlated with litter size in the flock（P 〈 0.05）, however, did not interact with mtDNA T7719G mutation（P 〉 0.05）.Conclusions： Our results highlight mutations of ovine mitochondrial coding genes, suggesting T7719G in tRNA-Lys gene be a potentially useful marker for selection of sheep litter size.
文摘旨在利用全基因组关联分析探寻敖汉细毛羊羊毛性状新的分子标记和候选基因。本研究采集1~2周岁的健康敖汉细毛羊耳组织与羊毛作为试验素材,其中,母羊248只,公羊81只,总计329只。羊毛进行性状测定(包括纤维直径、自然长度、伸直长度、伸直率),并对表型数据进行描述性统计和相关性分析。利用绵羊40K液相SNP芯片对全部个体进行基因分型。使用Plink 1.07软件对芯片数据进行质控,使用GCTA软件和PopLDdecay软件对质控数据进行群体结构分析。利用GMEMA混合线性模型对4种羊毛性状进行了全基因组关联分析(genomewide association study,GWAS),利用在线软件对候选基因进行GO和KEGG富集分析。质控后得到329只个体的30079个SNPs位点用于后续分析。通过GWAS分析筛选出4个在全基因组上显著相关的SNPs位点可能影响羊毛经济性状,分别位于1号、6号及8号染色体上。筛选出9个在染色体水平上显著相关的SNPs位点可能对羊毛性状具有潜在意义,分别位于3、5、8、11、18、21、22、25号染色体,寻找到39个可能影响羊毛性状的候选基因。本研究结果为后续探究敖汉细毛羊羊毛性状的遗传机制及分子育种标记开发提供重要参考。
文摘为了研究阿勒泰大尾羊和小尾寒羊不同脂肪组织及其他组织之间基因表达的差异,探讨脂肪与肥胖相关基因(fat mass and obesity associated gene,FTO)与绵羊体脂沉积的相关性。随机各选择6只6月龄雄性阿勒泰大尾羊和小尾寒羊,禁食24h后屠宰,分别采集下丘脑、海马、背最长肌、心肌和脂肪组织样本,应用冰冻组织切片技术,测定脂肪细胞的面积,用real-time PCR检测FTO基因在各组织中的表达水平。结果显示,阿勒泰大尾羊尾脂脂肪细胞面积极显著高于肾周脂脂肪细胞面积(P<0.01)。阿勒泰大尾羊与小尾寒羊下丘脑和海马FTO基因表达水平无显著差异,背最长肌、心肌FTO基因表达水平阿勒泰大尾羊的极显著高于小尾寒羊的(P<0.01),肾周脂中FTO基因表达水平阿勒泰大尾羊的显著高于小尾寒羊的(P<0.05)。阿勒泰大尾羊肾周脂、心周脂和尾脂中FTO基因表达水平在肾周脂、心周脂、尾脂之间无显著差异。结果表明,FTO基因表达水平在阿勒泰大尾羊和小尾寒羊之间存在品种间的差异。
文摘本研究旨在利用全基因组关联分析方法(genome-wide association studies,GWAS)挖掘影响绵羊体重性状的遗传标记和功能基因。采用Illumina OvineSNP50BeadChip中密度商业化羊芯片,对3个品种(苏尼特羊、德国肉用美利奴羊和杜泊羊)共计329只绵羊的体重性状(初生重、断奶重、6月龄重、断奶前日增重、断奶后日增重、日增重)进行GWAS分析。统计分析和基因注释基于TASSEL软件、混合线性模型和2012年10月公布的最新版绵羊基因组Ovis_aries_v3.1序列信息。结果表明,有10个SNPs位点在全基因组显著水平上与断奶后日增重相关,部分位点分布在羊注释基因内(MEF2B、RFXANK等);除此之外还检测到22个SNPs在染色体显著水平上与其他体重性状存在相关性,获得一批重要的候选基因。这些基因对绵羊体重性状功能基因的挖掘具有重要的理论意义和参考价值。
