湖羊、东湖杂种羊POMC基因外显子3单核苷酸多态性及其与生长性状的关联分析

阿黑皮素原(Pro-opiomelanocortin, POMC)在动物采食和能量平衡调控中发挥重要作用, 文章对绵羊POMC基因外显子3进行扩增和测序, 筛选多态性位点, 并分析多态位点与湖羊和东弗里生×湖羊杂种羊生长性状的相关性。测序后发现湖羊POMC基因外显子3有2个单碱基突变(g.273 T/C和g.456 G/A), 根据273位点处发生的T/C突变, 建立PCR-RFLP分析方法, 并对162只湖羊和130只东湖杂种羊进行检测分析。结果发现, 在湖羊群体中检测到TT(0.469)、TC(0.438)和CC(0.093)3种基因型, 而在东湖杂种羊群体中仅检测到TT(0.754)和TC(0.246)两种基因型。POMC基因外显子3的273位点多态性与生长性状的相关性研究结果显示：湖羊群体中CC基因型个体的2月龄断奶重、4月龄尻高及TC基因型个体4月龄体长和管围均显著高于TT型个体(P<0.05); CC基因型个体的4月龄重、6月龄重极显著高于TT和TC基因型个体(P<0.01); CC基因型个体的4月龄体高和体长极显著高于TT型个体(P<0.01), 且显著高于TC基因型个体(P<0.05)。此外, CC型个体的管围极显著高于TT基因型个体(P<0.01)。东湖杂种羊群体中TC基因型个体的2月龄断奶重、4月龄重及4月龄体高、体长、胸深和管围都显著高于TT型个体(P<0.05), TC型个体的6月龄重极显著高于TT型个体(P<0.01)。研究结果表明, POMC基因外显子3与绵羊生长性状相关, C等位基因对体重及体尺性状的增加更有利。该结果为进一步探讨POMC基因作为绵羊生长性状的辅助选育标记奠定了基础。     

单胺氧化酶B基因单核苷酸多态性与藏獒对陌生人攻击行为的关联分析

采用行为测试的方法,测定藏獒和拉布拉多犬在陌生人接近时的攻击行为差异,应用PCR-RFLP技术检测单胺氧化酶B(MAOB)基因第3外显子单核苷酸多态性,分析其在藏獒(陌生人接近时进行攻击)和拉布拉多犬(陌生人接近时无攻击行为)两个品种间基因型频率与基因频率分布差异.结果 表明:MAOB基因型频率与基因频率在两个品种中分布差异不显著(P＞0.05).x2适合性检验显示,藏獒处于Hardy-Weinberg极不平衡状态(P＜0.01),拉布拉多犬处于Hardy-Weinberg平衡状态(P＞0.05).藏獒和拉布拉多犬品种间,在陌生人接近时的攻击行为差异与MAOB基因T119C多态性可能无直接关系.     

山羊GOLA-DQB1基因外显子2多态性与免疫性状的相关分析

利用PCR-RFLP技术, 对莱芜黑山羊、鲁波山羊和波尔山羊3个山羊种群共 175 只个体的GOLA-DQB1基因外显子2进行遗传多态性研究, 并对山羊种群的血液免疫指标的效应进行了分析。结果表明: 3个山羊种群共检测到(AA、BB、CC、AB、AC、BC、DD)7种基因型, GOLA-DQB1基因外显子2的第24、151位的碱基表现出多态性。多数指标品种效应是主要效应。莱芜黑山羊中, BC基因型的淋巴细胞百分比(W-SCR)显著高于AC 、CC基因型(P<0.05), 中性球比例(W-LCR)显著低于CC基因型(P<0.05), 大型白细胞数(W-LCC)低于AC、CC基因型, 但差异不显著(P>0.05)。波尔山羊中, BC基因型的W-LCC低于AA 、AB 、BB基因型, 但差异不显著(P>0.05)。鲁波山羊中, BC 、AC基因型的W-LCC显著低于AA基因型(P<0.05)。揭示GOLA-DQB1基因与血液免疫性状有一定的相关性。     

中华鳖GHRL基因SNPs的筛选及生长性状的关联分析

为探究GHRL基因多态性对中华鳖(Pelodiscus sinensis)生长性状的影响, 采用直接测序法在GHRL基因上检测到14个单核苷酸多态性位点C289T、G501T、T738C、G776T、A841G、T885C、T2960C、A2987T、G3390A、A3857C、G4718A、T4820C、A4850C、T4979C。随机选取同批繁殖的120只中华鳖用飞行时间质谱法进行SNPs位点的分型, 并分析与生长性状的相关性。检测结果显示, 所有SNP位点均符合Hardy-Weinberg平衡状态(P>0.05)。方差分析结果显示, C289T位点CT、CC基因型的5项生长数据均显著高于TT基因型(P<0.05)。S2位点AB基因型的体重、背甲长、背甲宽和裙边宽4项数据均显著高于AA基因型(P<0.05)。G3390A位点AG基因型的背甲长、背甲宽2项数据显著高于AA基因型(P<0.05)。G4718A位点AG基因型的背甲长、背甲宽、裙边宽3项数据显著高于AA基因型(P<0.05)。在GHRL基因上获得的SNP位点可能影响着中华鳖的生长性状或与之紧密连锁, 可为中华鳖分子辅助育种提供助力与参考。     

SIM1基因第8外显子的SNP对猪背膘厚的影响

采用PCR-SSCP方法检测了约克夏、杜洛克、皮特兰、长白猪、嘉兴黑猪和金华猪6个品种共169头猪的SIM1基因外显子8的SNP及其基因型频率。结果共发现CC、CT、TT 3种基因型, 其基因型频率在国内外猪品种之间具有较大差异。其中, 国内猪种嘉兴黑猪和金华猪只存在TT基因型, 而国外猪种约克夏、杜洛克、皮特兰、长白猪则都存在3种基因型。用最小二乘法分析SNP对长白猪、约克夏猪和杜洛克猪的背膘厚的效应的结果表明, 纯合基因型个体的背膘厚大于杂合基因型个体。SIM1基因型对国外猪种背膘厚有显著效应(P< 0.05), 并且不同部位效应不同。     

新疆维吾尔族人载脂蛋白A5-1131T＞C基因多态性对血脂水平的影响

目的:为了探讨新疆维吾尔族人ApoA5基因-1131T＞C基因变异的频率及对血脂水平的影响.方法:对145例经冠脉造影排除冠心病的维吾尔族患者,采用聚合酶链反应-限制性片段长度多态性分析对ApoA5基因-113IT＞C多态性进行检测,比较不同基因型与个体血脂水平的关系.结果:ApoA5-1131C等位基因的频率是33%.TT、TC和CC三种基因型中,TC型与CC型的血甘油三酯(TG)水平明显高于TT型,而以CC型的TG水平最高(P均＜0.05),C携带者(TC CC)较非C携带者(TT)的TG水平增高36.2%[(2.03 1.33)mmol/Lvs(1.49 1.06)mmol/L,P＜0.05].结论:维族人群的ApoA5-113IT＞C基因变异较常见,-1131C等位基因与血TG水平增高有关.     

载脂蛋白A5基因-1131 TC、-12238 TC位点SNP与男性冠心病及血脂水平的相关性

目的研究男性患者中载脂蛋白A5基因(apolipoprotein A5 gene,APOA5)-1131 TC、-12238 TC位点多态性与冠心病及血脂水平的关系。方法选取2013.1-2015.1期间我院收治的男性冠心病患者226例作为观察组,同期选择195例来我院体检的男性健康人作为对照组。均采用聚合酶联反应-限制性片段长度多态性分析,检测-1131 TC、-12238 TC位点基因多态性,并测量血脂水平,观察多态性与冠心病、血脂的关系。结果两组对象-1131TC、-12238 TC位点中TT基因型均无差异,P0.05;而TC、CC基因型均有差异,P0.05;-1131 TC、-12238 TC位点中TC基因型与TT、CC基因型的TG、TC水平均有差异,P0.05;而HDL-C、LDL-C水平各基因型均无差异,P0.05。结论 TC、CC基因型与男性冠心病具有一定关系,而TC基因型对血脂TG、TC有明显影响,可能是引起冠心病的原因之一。     

大白猪BF基因多态性与繁殖性状及胎盘效率关联研究

文章以BF基因为产仔数的候选基因, 运用PCR-RFLP方法对大白猪BF基因内含子1序列进行分析, 发现内含子1第79 bp处发生了C→T突变, 命名为BF-intron 1-C79T。所扩增PCR产物经SmaⅠ酶切后, 可分出3种基因型, 分别为CC、CT和TT。经χ²适合性检验, 大白猪群体在该位点达到了Hardy-Weinberg平衡状态。将不同基因型与总产仔数(TNB)、产活仔数(NBA)、初生重(BW)和胎盘效率(PE)进行关联分析, 结果发现, 初产母猪在该位点上, CC基因型比CT基因型个体的TNB和NBA均多3.10头, 但差异不显著(P>0.05); 在BW和PE上, CC基因型均高于CT基因型, 差异不显著(P>0.05)。经产母猪在该位点上, CC基因型比TT基因型个体的TNB和NBA分别多3.45头和3.92头, PE高23.80%, 均达到显著水平; 但CT基因型个体与CC和TT个体之间的TNB 、NBA和PE差异不显著(P>0.05)。因此, BF-intron 1-C79T位点可作为繁殖性状及胎盘效率的潜在分子育种标记, 具有很大的研究价值。     

早胜牛类群A-FABP基因SNPs及其与胴体品质和肉质性状的相关性分析

研究早胜牛类群A-FABP基因多态性及其与胴体品质和肉质性状的相关性。采用PCR-SSCP方法对5个早胜牛类群(庆阳类群、平凉类群、南德温与庆阳类群杂种、西门塔尔与平凉类群杂种、秦川牛与平凉类群杂种)的A-FABP基因进行多态性分析,分析基因型与胴体品质和肉质性状的相关性。结果显示,A-FABP基因第三外显子区存在c.408GC的突变,并检测到3种基因型GG、GC和CC。胴体性状相关分析表明,GG基因型的胴体重显著低于CC基因型(P0.05);GG基因型屠宰率极显著低于GC基因型(P0.01),显著低于CC基因型(P0.05);GG基因型净肉率显著低于GC基因型(P0.05);GG基因型的眼肌面积显著低于GC和CC基因型(P0.05)。肉质性状相关分析表明,GG基因型失水率显著高于CC基因型个体(P0.05);GG基因型剪切力极显著高于GC和CC基因型(P0.01);GG基因型蒸煮损失和pH均显著高于GC基因型(P0.05),极显著高于CC基因型(P0.01)。A-FABP基因突变位点可作为胴体性状和肉质性状遗传标记。     

兴义鸭LPL基因外显子8多态性与生长和肉质性状的关联性分析

脂蛋白脂酶(LPL)是动物脂质沉积和新陈代谢的关键酶,对生长和肉质发挥着重要作用。试验采用PCR-SSCP法和DNA直接测序技术相结合对兴义鸭L PL基因外显子8进行多态性检测,并分析其多态与生长和肉质性状的关联性。结果表明,在兴义鸭L PL基因外显子8首次检测到1个T1251C同义突变,产生三种基因型TT、TC和CC,2个等位基因T和C,基因型TT和等位基因T的频率分别为0.8077和0.8750,多态信息含量为0.1948,表现为低度多态,卡方(字2)检验表明T1251C位点的基因型分布在兴义鸭中未偏离Hardy-Weinberg平衡。最小二乘分析显示,TT基因型个体的宰前体重显著低于CC基因型(p<0.05),胸宽显著低于TC基因型(p<0.05),推测等位基因C可能是兴义鸭生长性状的有利等位基因,可作为生长性状的一个标记性辅助选择位点。     

Polymorphisms of <Emphasis Type="Italic">STAT5A</Emphasis> gene and their association with milk production traits in Holstein cows

The STAT5A gene was studied as a candidate gene for five milk production traits (milk yield at 305 days, protein percentage, fat percentage, lactose percentage and dry matter percentage) in Holstein cows. According to the sequence of bovine STAT5A gene, two pairs of primers (P1 and P2) were designed to detect polymorphisms of STAT5A gene in 401 Holstein cows by PCR-RFLP and PCR-SSCP. The results showed that the products amplified by primers P1 and P2 displayed polymorphisms. For P1, three genotypes (AA, AG, and GG) were detected, and the frequency of AA/AG/GG was 0.252/0.486/0.262, respectively. Sequence analysis revealed a single nucleotide substitution A–G at 14217 bp (GenBank NC_007317) of bovine STAT5A gene while compared GG genotype with AA genotype. The differences of the least squares means for the four milk production traits (milk yield at 305 days, fat percentage, lactose percentage and dry matter percentage) between AA, AG and GG were not significant (P > 0.05). Least squares mean of protein percentage for AG or GG was significantly higher than that for AA (P < 0.05); the difference of the least squares mean for protein percentage was not significant between AG and GG (P > 0.05). For P2, three genotypes (CC, CT, and TT) were detected in Holstein cows, and the frequency of CC/CT/TT was 0.751/0.234/0.015, respectively. Sequencing revealed an insertion CCT at 17266 (NC_007317) of bovine STAT5A gene while compared CC genotype with TT genotype. The differences of the least squares means for the three milk production traits (protein percentage, lactose percentage and dry matter percentage) between CC, CT and TT were not significant (P > 0.05). Least squares mean of milk yield at 305 days for TT or CT was significantly higher than that for CC (P < 0.05); the difference of the least squares mean for milk yield at 305 days was not significant between TT and CT (P > 0.05). Least squares mean of fat percentage for CC or CT was significantly higher than that for TT (P < 0.05); the difference of the least squares mean for fat percentage was not significant between CC and CT (P > 0.05). The results preliminarily indicated that allele G of A14217G polymorphic site of STAT5A gene is a potential DNA marker for improving protein percentage in dairy cattle, 17266indelCCT polymorphic site of STAT5A gene is a potential DNA marker for improving milk yield at 305 days and fat percentage in dairy cattle.     

荷斯坦牛HSP70-1基因遗传多态性与乳腺炎抗性关系

以253头中国荷斯坦奶牛为研究对象, 检测HSP70-1基因的多态性, 并分析其多态性与中国荷斯坦牛体细胞评分(Somatic cell score, SCS)的相关性。首先以PCR-SSCP法寻找HSP70-1基因编码区的突变, 并通过测序确定突变的类型, 根据突变类型寻找合适的内切酶, 最终采用PCR-RFLP方法鉴定实验牛基因型; 然后分析基因多态性与中国荷斯坦牛SCS的相关性。结果表明HSP70-1基因的1 623 bp处产生G→A→C突变, 2 409 bp处产生G→A突变, 两位点都是沉默突变, 未引起氨基酸序列的改变; 经χ2 适合性检验, 中国荷斯坦牛在两个位点均未达到Hardy-Weinberg平衡状态; 同时, 群体基因座不同基因型与SCS相关分析的结果表明, 2409位点基因型与SCS相关性不显著(P>0.05), 1623位点基因型与SCS相关性显著(P<0.05), CC型SCS显著低于AG、GG型(P<0.05), CC基因型为乳腺炎抗性基因型。在中国荷斯坦奶牛群体中, HSP70-1基因CC基因型可作为改良奶牛乳腺炎抗性性状的分子遗传标记。     

MTHFR C677T、A1298C基因多态性与老年单纯收缩期高血压患者Hcy、血脂水平的关系研究

摘要 目的：研究5,10-亚甲基四氢叶酸还原酶（MTHFR）C677T、A1298C基因多态性与老年单纯收缩期高血压（ISH）患者同型半胱氨酸（Hcy）、血脂水平的关系。方法：选取2019年3月至2021年3月期间中南大学湘雅医学院附属海口医院全科医学科收治的212例老年ISH患者作为ISH组,以同期体检无高血压老年人120例为对照组。检测两组MTHFR C677T、A1298C基因多态性。收集两组一般资料及血浆Hcy及血脂检查结果。观察MTHFR C677T、A1298C不同基因型的血浆Hcy、血脂水平差异。采用多因素Logistic回归分析老年ISH发生的影响因素。结果：相比于对照组,ISH组MTHFR C677T位点T等位基因频率较高,C等位基因频率较低;ISH组CC基因型频率较低,CT、TT基因型频率较高（P<0.05）。相比于对照组,ISH组A1298C位点C等位基因频率较高,A等位基因频率较低;ISH组A1298C位点AA基因型频率较低,CC、AC基因型频率较高（P<0.05）。MTHFR基因C677T位点不同基因型血浆Hcy、总胆固醇（TC）水平差异具有显著性（P<0.05）。MTHFR基因A1298C位点不同基因型血浆Hcy、TC水平明显差异具有显著性（P<0.05）。血浆Hcy、MTHFR C677T及A1298C基因多态性是老年ISH发生的影响因素（均P<0.05）。结论：MTHFR C677T、A1298C基因多态性与老年ISH患者血浆TC、Hcy水平有关,血浆Hcy、MTHFR C677T及A1298C基因多态性是老年ISH发生的影响因素。     

ERα-29位基因多态性与HBV相关原发性肝癌易感性的关系

目的:探讨ERα-29位基因多态性与HBV相关原发性肝癌(PHC)易感性的关系。方法:选择100例HBV相关原发性肝癌患者作为研究组,另选择100例健康体检者作为对照组。应用聚合酶链反应-限制性片段长度多态性法检测两组ERα-29位基因多态性。结果:研究组ERα-29位基因TT和TC基因型分布高于对照组,而CC基因型分布低于对照组,差异具有统计学意义(P0.05);研究组T等位基因频率高于对照组,而C等位基因频率低于对照组,差异具有统计学意义(P0.05)。T等位基因发生PHC的风险高于C等位基因(P0.05)。结论:ERα-29位基因多态性与HBV相关原发性肝癌的易感性有关,其中T等位基因发生PHC的风险较高。     

Oxidative stress mediates cardiac fibrosis by enhancing transforming growth factor-beta1 in hypertensive rats

The methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism has been shown to be associated with cardiovascular disease and in patients with end-stage renal disease (ESRD). However, the relationship between MTHFR polymorphisms and cardiovascular disease (CVD) in patients on hemodialysis has not been examined. The aim of this study was to assess the association of polymorphisms of MTHFR gene with homocysteine (Hcy) and intimal medial thickness (IMT) in patients on hemodialysis. We performed case-control study involving107 patients with ESRD and 103 healthy controls. Plasma Hcy was measured in all the subjects and these subjects were genotyped for three MTHFR polymorphisms (C677T, A1298C, and G1793A). We observed significantly higher Hcy levels in patients as compared to controls. The frequency of MTHFR 1298CC genotype was significantly higher in ESRD patients than in controls (21.4% vs. 2.9%); the frequency of the MTHFR C677T genotypes did not differ between groups (26.1% vs. 17.4%). Compound heterozygous MTHFR 677CT/1298AC genotypes showed maximum association with the risk of ESRD (OR: 12.8; 5%CI: 1.64–10.01, P < 0.05). Concurrent occurrence of MTHFR 677CC wild genotype with either 1298CC or 1793GA significantly increased the risk of disease (OR: 7.20; 95%CI: 2.06–2.51, P < 0.001 and OR: 7.60; 95%CI: 1.68–34.35; P < 0.05, respectively). MTHFR 1298CC genotype was associated with higher Hcy levels. IMT was also significantly higher in patients with the 1298CC genotype (P < 0.05). Thus, A1298C polymorphism of MTHFR gene appears to be associated with the severity of carotid atherosclerosis and co-occurrence of MTHFR polymorphisms may be a risk factor for CVD in patients on hemodialysis.     

Polymorphisms of BMPR-IB gene and their relationship with litter size in goats

The bone morphogenetic protein receptor IB (BMPR-IB) gene was studied as a candidate gene for the prolificacy of goats. According to mRNA sequence of ovine BMPR-IB gene, ten pairs of primers were designed to detect single nucleotide polymorphisms (SNPs) of exon 1, exon 2, exon 6 to exon 10 and 3′ untranslated region (UTR) of the BMPR-IB gene in both high prolificacy breed (Jining Grey goat) and low prolificacy breeds (Wendeng Dairy and Inner Mongolia Cashmere goats) by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) method. Only the products amplified by primers P8 and P10 of the 3′UTR displayed polymorphisms. For primer P8, three genotypes (AA, AB and BB) were detected in Jining Grey and Wendeng Dairy goats, two genotypes (AA and AB) were in Inner Mongolia Cashmere goats. Sequencing revealed one mutation (71C→T) of the BMPR-IB gene in genotype BB compared with AA. The differences of least squares mean (LSM) for litter size between genotypes AA, AB and BB were non-significant (P > 0.05) in Jining Grey goats. For primer P10, three genotypes (CC, CD and DD) were detected in Jining Grey and Wendeng Dairy goats and one genotype (CC) in Inner Mongolia Cashmere goats. Sequencing revealed one mutation (130T→C) of the BMPR-IB gene in genotype DD compared with CC. The differences of LSM for litter size between genotypes CC, CD and DD were non-significant (P > 0.05) in Jining Grey goats. These results preliminarily showed that the detected loci of the BMPR-IB gene had no significant effect on prolificacy of Jining Grey goats.     

A novel SNP of liver-type fatty acid-binding protein gene in duck and its associations with the intramuscular fat

Liver-type fatty acid-binding protein (L-FABP) is a member of intracellular lipid-binding proteins involved in the transportation of fatty acids. We detected the polymorphisms of duck L-FABP gene and its association with the intramuscular fat (IMF) and other fat-related traits. The complete sequence of duck L-FABP gene (four exons and three introns, 2,542?bp) was obtained in this study. The polymorphism of L-FABP gene was examined with direct DNA sequencing method in 231 individuals from different breeds, and a novel single nucleotide polymorphism in the exon 3 was detected. The polymorphism was shown to be associated with the contents of C16:0, C18:3 and the total IMF in pectoral muscle. The content of C16:0 in genotype CC was significantly higher than CT (P?<?0.01) and TT (P?<?0.01), and the genotype CT was higher than TT (P?<?0.01). The content of C18:3 in genotype TT was significantly higher than CC and CT (P?<?0.01), whereas the genotype CC and CT had no significant difference (P?>?0.05). The content of IMF in genotype CC was significantly higher than CT (P?<?0.01). However, no significant difference was detected between genotype CC and TT or genotype CT and TT (P?>?0.05).     

牛生长激素受体基因多态性与体尺、体重指标的相关性分析

利用PCR-SSCP技术对南阳牛、利木赞、盖洛威共100头牛的生长激素受体基因的外显子10(GHR10)部分序列进行单核苷酸多态性研究, 并分析了该基因的不同基因型与3个品种牛生产性状的关系。结果表明: 南阳牛、利木赞、盖洛威3个品种中共存在6种基因型(AA、BB、CC、AB、AC、BC), c2检验表明该实验群体在这一位点上处于Hardy-Weinberg平衡状态(P > 0.05), 利木赞和盖洛威的PIC表现为中度多态,而南阳牛表现高度多态。测序结果显示: 所扩增GHR10部分片段共有5处碱基突变, 分别是495 bp (A/T), 622 bp(C/T), 650 bp(A/C), 702 bp(T/C), 730 bp(A/G); 并导致3处氨基酸替代: 其中622 bp Pro/Ser(脯氨酸/丝氨酸), 650 bp Asn/Thr(天冬酰胺/苏氨酸), 730 bp Ser/Gly(丝氨酸/甘氨酸)。最小二乘分析表明: 基因型AB、BC所对应的12月龄体重最小二乘均值显著高于基因型CC所对应的最小二乘均值(P < 0.05, P < 0.01); 18月龄体重BC基因型显著高于CC基因型(P < 0.01), 基因型AB所对应的18月龄胸围最小二乘均值显著高于基因型CC所对应的最小二乘均值(P < 0.05)。