猪MSTN基因的多态性和生长性状关联分析

基于肌肉生长抑制素(Myostatin,MSTN)基因结构及功能研究报道,将其作为猪生长发育性状的候选基因进行多态性检测与关联分析,旨在为猪分子遗传标记提供依据。本研究以长白猪、大白猪、杜长大、通城猪、莱芜猪、五指山猪6个不同猪种基因组DNA为模板,通过克隆测序鉴定MSTN基因启动子区、第1内含子区、第2内含子区、第1外显子区、第2外显子区及3′UTR区SNPs位点;以长白猪和杜长大2个试验猪群为材料,利用基质辅助激光解吸飞行时间质谱法对MSTN基因进行SNP分型。建立最小二乘法分析模型,利用SAS软件分析数据。结果表明:在6个猪种76个个体中,共筛选出16个SNPs,其中有4个位于启动子区,5个位于第1内含子,7个位于第2内含子,在外显子1、外显子2和3′UTR区域并未检测到SNP位点。对MSTN基因的4个SNPs位点(P1、P3、P4、P5;其中P1、P3、P4位于启动子区,P5位于内含子1区)的生长性状关联分析显示,P4和P5 2个位点的多态性与猪生长性状显著相关(P<0.05)。P4和P5 2个位点具有作为分子标记辅助猪育种的潜在应用价值。     

东佛里生羊MSTN基因多态性及其与生长性状的关联分析

试验旨在研究肌肉生长抑制素(myostatin,MSTN)基因多态性及其对东佛里生羊生长性状的影响。从324只东佛里生羊全血中提取DNA,对MSTN基因外显子序列和非翻译区(untranslated region,UTR)序列进行PCR扩增,采用一代测序技术鉴定单核苷酸多态性(single nucleotide polymorphism,SNP)位点,分析不同位点的遗传多态性,并将不同基因型与东佛里生羊的生长性状进行关联分析。结果显示,在东佛里生羊MSTN基因外显子中未检测到突变;在3'-UTR区的272 bp处存在G→A单碱基突变,该位点只检测到AG和AA基因型,且AA基因型频率高于AG基因型;在5'-UTR区的176 bp处存在C→A突变,该位点只检测到CC和AC基因型,且CC基因型频率高于AC基因型。遗传多样性参数分析结果表明,东佛里生羊群体MSTN基因3'-UTR-272和5'-UTR-176位点的纯合度分别为0.827和0.887,杂合度分别为0.173和0.113,2个位点的多态性均较低(PIC<0.25),其群体基因型分布均符合哈代-温伯格平衡(P>0.05)。关联分析发现,3'-UTR-272位点AG基因型体斜长显著高于AA基因型(P<0.05),5'-UTR-176位点CC基因型胸围显著高于AC基因型(P<0.05),其余指标均无显著差异(P>0.05)。综上,东佛里生羊MSTN基因3'-UTR和5'-UTR突变位点可作为其目标性状选育的分子标记。     

不同绵羊品种GHSR基因多态性与生长性状的关联分析

以GHSR基因为候选基因,随机选取滩羊、小尾寒羊和兰州大尾羊3个品种共计1 249个个体为试验材料,利用DNA池测序结合琼脂糖凝胶电泳方法,检测突变多态性,分析其与体重、生长性状的相关性。试验结果表明,在GHSR基因的第一内含子上存在24 bp的插入/缺失突变,3个绵羊品种中均存在AA、AB、BB 3种基因型。该位点的多态性与3种绵羊8个生长性状指标关联分析显示,兰州大尾羊BB基因型的个体体重显著高于其他基因型;AB基因型的个体体高、十字部高显著高于其他基因型;小尾寒羊AB基因型的个体胸宽显著高于其他基因型;AA基因型的个体体高、十字部高显著高于BB基因型;滩羊AB基因型的个体体斜长、体重显著高于AA基因型,BB基因型的个体体高显著低于其他基因型。研究表明:GHSR基因可以作为3种绵羊生长性状的候选基因,为绵羊育种选育提供新思路。     

绵羊MSTN基因多态性及其与肉质性状的关联分析

【目的】探讨绵羊肌肉生长抑制素(myostatin, MSTN)基因多态性及其对绵羊肉质性状的影响,以期为绵羊肉用性能的选育提供有效的遗传分子标记。【方法】利用液相捕获测序技术获取MSTN基因在杜泊羊(D)、滩羊(T)和小尾寒羊(XH)共91只绵羊中的变异位点,并筛选出品种间差异显著的单核苷酸多态性(single nucleotide polymorphism, SNP)位点。针对筛选出的位点,利用飞行质谱检测技术对滩羊、小尾寒羊、杜泊羊和杜滩寒三元杂交羊(DTH)共30只绵羊进行基因分型,并与肉质性状进行关联分析。【结果】试验筛选出的MSTN基因rs129059715位点在4个绵羊群体中共检测到3种基因型,分别为15个CC基因型、9个CA基因型及6个AA基因型,C为优势等位基因(除杜滩寒三元杂交羊外)。在杜泊羊、滩羊和杜滩寒三元杂交羊群体中均处于Hardy-Weinberg平衡状态(P>0.05),在小尾寒羊群体中处于Hardy-Weinberg不平衡状态(P<0.05);在杜泊羊和小尾寒羊群体中均处于低度多态(PIC<0.25);在滩羊和杜滩寒三元杂交羊群体中均处...     

绵羊FUT8基因多态性及其与生长性状的关联分析

【目的】研究岩藻糖基转移酶8(fucosyltransferase 8,FUT8)基因g.74522417 C>T位点多态性及其对绵羊生长性状的影响,为绵羊分子育种筛选新的遗传标记。【方法】采用Sequenom MassARRAY~? SNP分型技术、Illumina OvineSNP 600K BeadChip及Illumina OvineSNP 50K BeadChip芯片检测技术对湖羊(n=992)和乌珠穆沁羊群体(n=333)FUT8基因g.74522417 C>T位点进行分型,并将其多态性与湖羊和乌珠穆沁羊的生长性状进行关联分析。【结果】FUT8基因g.74522417 C>T位点在乌珠穆沁羊和湖羊群体中均检测出3种基因型：CC、TC和TT。群体遗传分析表明,g.74522417 C>T位点在乌珠穆沁羊和湖羊群体中均处于中度多态(0.25T位点在乌珠穆沁羊和湖羊群体中均处于哈代-温伯格平衡状态(P>0.05)。关联分析结果表明,FUT8基因g.7452241...     

绵羊IGF1R基因多态性及其与生长性状的关联分析

【目的】探究胰岛素样生长因子1受体(insulin-like growth factor 1 receptor, IGF1R)基因遗传变异对绵羊生长性状的影响,以期为优质肉羊新品种(系)培育提供有效的分子遗传标记。【方法】利用液相捕获测序技术获得IGF1R基因在杜泊羊、小尾寒羊和滩羊共91只羊中的变异位点,筛选出品种间差异显著的单核苷酸多态性(single nucleotide polymorphism, SNP)位点。针对筛选出的位点利用飞行质谱对3个绵羊品种杂交后代383只绵羊进行检测,并与初生及3月龄绵羊的生长性状进行关联分析。【结果】3个绵羊品种IGF1R基因共检测到347个突变位点,其中外显子SNPs共8个：g.7628315 T>C、g.7628528 T>C、g.7628690 C>T、g.7833817 C>T、g.7836687 C>T、g.7840691 T>C、g.7855904 C>G、g.7872277 C>T,在群体中均表现出多态性,χ²检验结果表明,除小尾寒羊g.7628690 C>...     

牦牛MDHⅠ基因多态性及其与生长性状的关联分析

本研究旨在探讨牦牛细胞质苹果酸脱氢酶（cytoplasmic malate dehydrogenase,MDHⅠ）基因的遗传多态性及其与生长性状的关联性,以期为牦牛优良性状选育提供参考依据。选取5个牦牛类群（共178头）,采集耳组织样提取DNA并构建DNA池,对MDHⅠ基因所有外显子设计特异性引物进行扩增测序,用Mega 5.2筛查SNPs位点,直接测序法鉴定其基因型,利用PopGene 32进行χ²独立性检测、基因纯合度（Ho）、基因杂合度（He）、有效等位基因频率（Ne）和多态信息含量（PIC）分析,利用SPSS 24.0分析MDHⅠ基因与牦牛生长性状间的关联性。结果表明,申扎牦牛、类乌齐牦牛、斯布牦牛、帕里牦牛和麦洼牦牛均在外显子8区域发现1个突变位点（G23093C）,属同义突变,有3种基因型：GC、CC和GG,5个牦牛类群中,GC基因型为优势基因型（类乌齐牦牛中GG为优势基因型）,G为优势等位基因（斯布牦牛除外）,在斯布牦牛中,G和C基因频率相等。χ²适应性检验表明,申扎牦牛、类乌齐牦牛、斯布牦牛、帕里牦牛和麦洼牦牛均符合Hardy-Weinberg平衡（P>0.05）。MDHⅠ基因在申扎牦牛、帕里牦牛和斯布牦牛中属于高度多态（PIC>0.5）,在麦洼牦牛和类乌齐牦牛中属于中度多态（0.25 < PIC < 0.5）;纯合度最高的是申扎牦牛和类乌齐牦牛。关联性分析结果表明,MDHⅠ基因不同基因型对类乌齐牦牛、帕里牦牛和麦洼牦牛体重有显著性影响（P<0.05）;且种间分析发现,MDHⅠ基因不同基因型对牦牛体重有显著性影响（P<0.05）,说明该基因可作为人工选育的分子标记。     

奶山羊AGPAT6基因多态性与生长性状的关联分析

利用PCR-RFLP技术对549只西农萨能和关中奶山羊群的AGPAT6基因遗传多态性与生长性状(体高、体长和胸围)的相关性进行了研究。结果表明AGPAT6基因存在4个SNPs多态:P1、P2、P4与P10位点分别位于5'UTR(g.152G>C)、内含子2(g.8124G>A)、外显子4(g.9263C>G)和3'UTR(g.16436G>A)。经关联分析,P2、P4和P10三位点在西农萨能奶山羊和关中奶山羊品种中都属于中度多态,4个SNPs的不同基因型对两个奶山羊品种的生长性状影响均不显著(P>0.05)。     

奶山羊AGPAT6基因多态性与生长性状的关联分析

利用限制性片段长度多态性聚合酶链反应技术对549只西农萨能和关中奶山羊群的AGPAT6基因遗传多态性与生长性状(体高、体长和胸围)的相关性进行研究.结果表明AGPAT6基因存在4个单核苷酸多态性:P1、P2、P4与P10位点分别位于5′UTR (g.152G＞C)、内含子2(g.8124G＞A)、外显子4(g.9263C＞G)和3′UTR(g.16436G＞A).经关联分析,P2、P4和P10三位点在西农萨能奶山羊和关中奶山羊品种中都属于中度多态,4个SNPs的不同基因型对两个奶山羊品种的生长性状影响均不显著(P＞0.05).     

绵羊生长性状间的相关和回归分析及TRHDE基因多态性与生长性状的关联分析

旨在探究乌珠穆沁羊生长性状间的相关性,构建体尺对体重的最优回归方程,分析TRHDE基因多态性对乌珠穆沁羊生长性状的影响。本研究选取336只生长发育良好的乌珠穆沁羊(公羊176只、母羊160只)作为研究对象,收集4月龄和6月龄生长性状,利用绵羊Illumina OvineSNP600 BeadChip在336只乌珠穆沁羊群体中对TRHDE基因SNPs进行基因型检测,采用SPSS 26.0软件对生长性状的相关性和回归性进行分析以及TRHDE基因多态性与生长性状进行关联分析。结果发现,乌珠穆沁羊4月龄和6月龄的公羊和母羊生长性状最优回归方程分别为：Y₁=-53.66+0.77X₁+0.39X₂、Y₂=-47.83+0.63X₁+0.45X₂、Y₃=-47.01+0.42X₁+0.13X₂+2.82X₃+0.19X₄+0.28X₅、Y...     

Polymorphism of CLPG and MSTN Genes and their Association Analysis with Growth Traits in Sheep

The study was conducted to explore the possibility that CLPG (Callipyge) and MSTN (Myostatin) genes which could be the candidate genes of sheep growth traits, and investigate the molecular genetic markers related to sheep growth traits.133 (Austrilian White sheep×Dorper sheep×Hu sheep) hybid-sheep were chosen as subjects, the technology of direct sequencing of PCR products and PCR-RFLP were used to detect the single nucleotide polymorphism of CLPG and MSTN genes, then the association of the SNPs different genotypes and combined genotypes with sheep growth traits were analyzed by the GLM statistical model of SPSS 22.0.Sequencing results showed that the SNP of C/T which called C1 was detected at position 232 bp of the STS sequence in CLPG gene.The SNP of G/A which called M1 was detected in the 3'UTR of MSTN gene.PCR-RFLP analysis showed that two genotypes CC and CT were in C1 site, two genotypes GG and GA were in M1 site.Association analysis revealed that C1 site was significantly or extremely significantly associated with backfat thickness and loin muscle area (P<0.05;P<0.01), M1 site was significantly or extremely significantly associated with body weight, tube girth, backfat thickness and loin muscle area (P<0.05;P<0.01).Meanwhile, the combined genotype was extremely significantly associated with body weight, backfat thickness and loin muscle area (P<0.01).The conclusions indicated that SNPs and combined genotype of CLPG and MSTN genes had effects on growth traits in sheep.C1 and M1 sites could be considered as effective genetic markers for sheep growth traits.     

澳洲白绵羊美丽臀和肌肉生长抑制素基因基因型的检测

美丽臀(callipyge,CLPG)及肌肉生长抑制素(myostatin,MSTN)是全世界公认的与肌肉生长发育相关的基因,突变后分别产生“美丽臀”和“双肌”的优良性状。为研究国外进口品种澳洲白绵羊是否携带2种基因的突变位点,本试验以澳洲白绵羊为试验动物,采用PCR-RFLP技术对CLPG、MSTN基因的多态性进行分型。结果发现:CLPG基因上有1个C→T的SNP位点,澳洲白绵羊群体中有CC和CT 2种基因型;MSTN基因在3'-UTR区发现1个G→A的SNP位点,澳洲白绵羊群体中有GG和GA 2种基因型。用R语言程序对所测数据进行性别间不同基因型与生长发育状况的单因素方差分析得知,CLPG和MSTN基因不同基因型对澳洲白公羊体重、体高、体斜长、尻宽、背膘厚和眼肌面积均无显著性影响(P>0.05);CLPG基因CT基因型在母羊体重上显著高于CC基因型(P<0.05),在体斜长上极显著高于CC基因型(P<0.01),MSTN基因GA基因型母羊眼肌面积极显著高于GG基因型(P<0.01)。澳大利亚进口澳洲白绵羊中筛选到含有CLPG和MSTN基因的突变位点,且该突变位点对绵羊体尺性状有显著性作用,因此澳洲白绵羊引进后可以作为改良本地羊品种、基因聚合及三元配套系的终端父本。     

绵羊FRZB基因InDel检测及其与生长性状的关联分析

本研究以卷曲相关同源蛋白（FRZB）基因作为候选基因,以同羊、小尾寒羊、兰州大尾羊和湖羊4个品种共计582只绵羊个体为试验材料,利用琼脂糖凝胶电泳结合DNA测序技术对FRZB基因的InDel进行筛查,旨在寻找与4个品种绵羊生长性状相关的InDel位点。结果表明,绵羊FRZB基因第1内含子上检测到一段14 bp的InDel突变;FRZB基因内含子区InDel突变位点在同羊、小尾寒羊、兰州大尾羊和湖羊群体中均存在II、ID、DD 3种基因型;关联分析表明,FRZB基因中检测到的InDel突变对同羊生长性状有显著影响（P<0.05）,其中II基因型的体长、背高、荐高、距骨宽度、尾长和尾宽在同羊群体中显著优于DD基因型,可以作为候选分子标记用于同羊分子标记辅助选择。这些结果提示,绵羊FRZB基因第1内含子上一个14 bp的插入突变可显著影响同羊生长性状,可以作为绵羊育种中生长性状的潜在分子标记。     

Expression of MSTN and MyoG Genes in Zi and Rhine Goose and their Correlation with Carcass Traits

The aim of this study was to investigate the effects of MSTN and MyoG genes on goose skeletal muscle growth.In this study,MSTN and MyoG genes expression were detected in breast and leg muscle of Zi and Rhine goose by Real-time PCR,and the correlations between genes expression levels and carcass traits were investigated.The results showed that the breast muscle weight and breast muscle rate of Rhine goose were extremely significant higher than Zi goose (P<0.01).MSTN and MyoG mRNA expression in breast muscle of Zi goose were significantly higher than that of Rhine goose,and the mRNA level of MSTN in leg muscle of Rhine was extremely significant higher than that of Zi goose (P<0.01),there was no significant difference of MyoG mRNA between Zi goose and Rhine goose (P>0.05).There was extremely significant difference between MSTN mRNA expression in breast muscle and leg muscle of Zi goose (P<0.01).MSTN mRNA expression in leg muscle was significantly higher than that of breast muscle of Rhine goose (P<0.05).There was extremely significant difference between MyoG mRNA expression in breast muscle and leg muscle of Zi goose and Rhine goose (P<0.01).There was a extremely significant negative correlation between MSTN mRNA expression in breast muscle with body weight,breast muscle weight and breast muscle percentage (P<0.01).There was a extremely significant positive correlation between MyoG mRNA expression in breast muscle and leg muscle with body weight (P<0.01).MSTN and MyoG gene might have positive and negative regulation effect on muscle growth.     

Polymorphism of MyoG Gene Exon 1 and Its Association Analysis with Meat Quality Traits in Sheep

Myogenin (MyoG) gene plays a central regulatory role in the process of muscle cell differentiation, which directly affects meat production capacity of animal. In this study, 6 sheep varieties including Large-tail Han sheep, Small-tail Han sheep, Yuxizhiwei sheep, Lanzhou fat-tailed sheep, Mongolia sheep and Tong sheep were selected as experimental materials to detect the polymorphism of MyoG gene exon 1 by non denaturing polyacrylamide gel electrophoresis, and analyze the association analysis between the polymorphism of MyoG gene exon 1 and meat quality traits in sheep. The results showed that 2 alleles (A, B) and 3 genotypes (AA, BB, AB) were detected in MyoG gene exon 1 of six sheep populations. The A allele frequency in Small-tail Han sheep, Large-tail Han sheep, Yuxizhiwei sheep, Lanzhou fat-tailed sheep, Mongolia sheep and Tong sheep were 0.5167, 0.2500, 0.4375, 0.6500, 0.5750 and 0.7125, respectively, the B allele frequency of six sheep populations were 0.4833, 0.7500, 0.5625, 0.3500, 0.4250 and 0.2875, respectively. MyoG gene exon 1 mainly affected the moisture and color of mutton, the moisture content of BB genotype was significantly higher than that of AB genotype (P< 0.01) and AA genotype (P< 0.05). The color of AB genotype was significantly higher than that of BB genotype (P< 0.01) and AA genotype (P< 0.05).     

绵羊MyoG基因外显子1的多态性及其与肉质性状的关联分析

肌细胞生成素（myogenin,MyoG）基因在肌细胞分化过程中起着中心调节作用,直接影响着动物的产肉能力。本研究以大尾寒羊、小尾寒羊、豫西脂尾羊、兰州大尾羊、蒙古羊、同羊6个绵羊品种为试验材料,利用非变性聚丙烯酰胺凝胶电泳检测MyoG基因外显子1的多态性,并与绵羊的肉质性状进行关联分析,探讨MyoG基因外显子1对绵羊肉质性状的影响。结果表明,MyoG基因外显子1在6个绵羊群体中均检测到3种基因型（AA、BB、AB）和2个等位基因（A、B）,在小尾寒羊、大尾寒羊、豫西脂尾羊、兰州大尾羊、蒙古羊、同羊群体中检测到MyoG基因外显子1的A等位基因频率分别为：0.5167、0.2500、0.4375、0.6500、0.5750和0.7125,MyoG基因外显子1的B等位基因频率分别为：0.4833、0.7500、0.5625、0.3500、0.4250和0.2875。MyoG基因外显子1主要对羊肉的水分和色泽有影响,主要表现为BB基因型水分极显著高于AB基因型（P< 0.01）,显著高于AA基因型（P< 0.05）;AB基因型的色泽极显著高于BB基因型（P< 0.01）,显著高于AA基因型（P< 0.05）。     

郏县红牛CLPG基因的SNP及对生长发育性状的影响

利用PCR-SSCP检测了113头郏县红牛CLPG基因的SNP，发现了AA、AB、AC 3种基因型，测序结果显示AA型在第150和419碱基处分别表现为C和T，为野生型；AB型在第419处发生碱基T→C突变；AC型在第150处发生碱基C→T突变。多态信息含量为0．226，属低度多态。分析了不同基因型与部分生长发育性状的相关性，结果显示，AA、AC基因型个体在体重、体长指标上显著（P〈0．05）或极显著（P〈0．01）大于AB型个体；在管围指标上，AA基因型个体优于AB基因型个体，差异显著（P〈0．05）；在坐骨端宽指标上，AA与AB基因型个体之间差异显著（P〈0．05），AB与AC基因型个体间差异极显著（P〈0．01）。     

Different Months of MSTN Gene mRNA Expression Levels and their Correlation with Growth Traits in Five Xinjiang Local Sheep

To explore the different varieties of five Xinjiang local sheep myostatin (MSTN) expression levels and their correlation with growth traits (body weight,body height,body length,heart girth and cannon circumference).The MSTN mRNA expression level in different month(Birth,1 months,2 months,3 months,4 months,5 months) of muscle and fat in Altay sheep,Turpan Black sheep,Emil White sheep,Kazakh sheep,Bashbay sheep by quantitative Real-time PCR method,using SPSS 19.0 software to analyze the correlation with growth traits.The MSTN mRNA expression levels in muscle:The MSTN gene mRNA expression levels of Altay sheep,Kazakh sheep and Bashbay sheep at birth were lower than other months;The MSTN gene mRNA expression level of Emil White sheep in 3 months was significantly lower than other months (P<0.05);The MSTN gene mRNA expression levels of Altay sheep,Turpan Black sheep were the highest in 4 months;The MSTN gene mRNA expression level of Kazakh sheep was the lowest at birth,and was the highest in 5 months.The MSTN gene mRNA expression levels in fat:The MSTN gene mRNA expression level of Emil White sheep at birth was significantly lower than other months (P<0.05); The MSTN gene mRNA expression levels of Altay sheep,Emil White sheep and Bashbay sheep were the highest in 5 months,the MSTN gene mRNA expression level of Bashbay sheep in 5 months was significantly higher than other months (P<0.05);The MSTN gene mRNA expression level of Kazakh sheep was the highest in 2 month,it was significantly higher than 5 months (P<0.05).Correlation analysis revealed that the MSTN gene mRNA expression levels were mostly negatively correlated with growth traits.The MSTN gene mRNA expression levels in muscle and fat of Altay sheep were all negatively correlated with growth traits;The MSTN gene mRNA expression levels in muscle of Turpan Black sheep were negative correlated with body length and body height,others were positively;The MSTN gene mRNA expression levels in muscle of Emil White sheep were positive correlated with heart girth and cannon circumference;The MSTN gene mRNA expression levels in fat were positive correlated with body length,others were negatively correlation;The MSTN gene mRNA expression levels in muscle and fat of Kazakh sheep were positive correlated with body length,others were negatively correlation;The MSTN gene mRNA expression levels in muscle of Bashbay sheep were positive correlated with body length and cannon circumference,others were negatively correlation.The MSTN gene mRNA expression level was discrepancy in different month of different varieties,and was no fixed expression pattern;The MSTN gene mRNA expression levels were positively correlated with the part of the body size.The MSTN gene mRNA expression levels might have a direct relationship with the growth traits and bone growth.