| 摘要: |
| 【目的】探究桥粒蛋白家族4（desmoglein 4，DSG4）基因的组织表达谱及其单核苷酸多态性与羊毛性状的关系。【方法】采集成年甘肃高山细毛羊的心脏、肝脏、脾脏、肺脏、肾脏、背最长肌及1，30，60，90，180和270日龄体侧部皮肤组织，以β-actin基因为内参基因，采用实时荧光定量PCR法（RT-qPCR）检测DSG4的表达谱。采集甘肃高山细毛羊体侧部羊毛样品及血样，测定羊毛性状指标（平均纤维直径、平均纤维长度、平均纤维曲度、舒适指数、平均纤维强度、纤维直径标准差及纤维直径变异系数）；提取血液全基因组的DNA，PCR扩增DSG4基因第9，11，12外显子及其上下游各150 bp的序列，测序后分析单核苷酸（SNP）多态性，并分析其SNP多态性与羊毛性状的相关性。【结果】DSG4基因在皮肤中高表达，在其他6种组织中几乎不表达；随着试验羊日龄的增加，皮肤组织中DSG4基因表达量呈先上升后下降的趋势，在90和180日龄时表达量较高，显著高于其他日龄。测序分析发现了g.30346351 G>A、g.30346381 T>C、g.30346414 G>T、g.30347673 G>C和g.30353299 G>A等5个SNPs，其中 g.30346351 G>A、g.30346381 T>C和g.30346414 G>T位点位于外显子9中，g.30347673 G>C位点位于外显子9下游150 bp序列内，g.30353299 G>A位点位于外显子12中。每个SNP位点均有3种基因型，均属于中度多态，且均处于哈代 温伯格平衡状态（P＞0.05）。5个SNPs位点中，g.30346351 G>A及g.30353299 G>A分别与羊毛纤维直径变异系数和平均纤维长度显著相关（P＜0.05）。【结论】DSG4基因对甘肃高山细毛羊羊毛平均纤维长度存在显著影响，可以将其SNPs位点作为该品种毛用性状选育的遗传分子标记。 |
| 关键词: 甘肃高山细毛羊 DSG4基因 单核苷酸多态性 基因组织表达 羊毛性状 |
| DOI：10.13207/j.cnki.jnwafu.2025.12.001 |
| 分类号: |
| 基金项目:甘肃省重点研发计划项目（22YF7WA115）；甘肃农业大学伏羲青年基金项目（Gaufx-03Y04）；国家留学基金委国合项目（20221007）；甘肃省高端外国专家引进计划项目（23JDKA0010） |
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| Association analysis between gene single nucleotide polymorphisms of the desmoglein 4 and wool traits in Gansu alpine fine wool sheep |
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MA Longxia, WU Bohua, HE Zhaohua, SUN Hongxian, CHEN Zhanzhao, ZHAO Fangfang, WANG Jiqing, LIU Xiu, HAO Zhiyun, LI Mingna, LI Shaobin
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College of Animal Science and Technology/Gansu Key Laboratory of Herbivorous Animal Biotechnology,Gansu Agricultural University,Lanzhou，Gansu 730070,China
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| Abstract: |
| 【Objective】This research aimed to explore the tissue expression profile of desmoglein 4 (DSG4) genes and their single nucleotide polymorphisms in relation to wool traits.【Method】Samples of hearts,livers,spleens,lungs,kidneys,and longest dorsal muscle,as well as lateral skin tissues of sheep at 1,30,60,90,180 and 270 d of age were collected from adult Gansu alpine fine wool sheep.With β-actin as the internal reference gene,the expression profile of DSG4 was detected by real time fluorescence quantitative PCR (RT-qPCR).Wool samples from the side of the Gansu alpine fine wool sheep and blood samples were collected to determine wool traits (mean fiber diameter,mean fiber length,mean fiber curvature,comfort index,mean fiber strength,fiber diameter standard deviation and variation coefficient of fiber diameter).The genomic DNA of the whole blood was extracted,and PCR was used to amplify the 9th,11th,and 12th exons of DSG4 gene and its upstream and downstream sequences of 150 bp each.The amplified products were sequenced,and their single nucleotide polymorphisms (SNPs) were analyzed.Then,the correlation between the SNP polymorphisms and wool traits was analyzed.【Result】DSG4 gene was highly expressed in the skin and barely expressed in the other six tissues.As the age of the experimental sheep increased,the expression level of DSG4 in their skin tissue showed a trend of first increasing and then decreasing.The expression level was relatively high at 90 and 180 days of age,significantly higher than those at other ages.Sequencing analysis revealed five SNPs,including g.30346351 G>A,g.30346381 T>C,g.30346414 G>T,g.30347673 G>C,and g.30353299 G>A.Among them,the g.30346351 G>A,g.30346381 T > C,and g.30346414 G>T loci were located in exon 9,g.30347673 G>C locus was located within 150 bp downstream sequence of exon 9,and g.30353299 G>A locus was located in exon 12.Each SNP locus had three genotypes,all of which belonged to moderate polymorphism and were in Hardy-Weinberg equilibrium state (P>0.05).Among the 5 SNPs loci,g.30346351 G>A and g.30353299 G>A were significantly correlated with the coefficient of variation of wool fiber diameter and fiber length (P<0.05).【Conclusion】DSG4 gene has a significant impact on the Mean fiber length of Gansu alpine fine wool sheep,and its SNPs loci can be used as genetic molecular markers for the selection of wool traits in this breed. |
| Key words: Gansu alpine fine wool sheep DSG4 gene single nucleotide polymorphism gene organization expression wool characteristics |
