绵羊X染色体59571364与59912586位点在脂尾、瘦尾绵羊群体中的多态性检测及分析

为了探讨绵羊X染色体上两处SNP(59571364和59912586)与绵羊尾脂沉积性状的关系, 继而为利用分子标记辅助选择育种技术培育低脂绵羊品种提供依据, 文章以尾型极端差异的阿勒泰羊、湖羊、中国美利奴细毛羊以及萨福克羊为研究对象, 利用PCR-RFLP检测两位点在群体中的多态性, 并分析了两个SNP位点在阿勒泰羊群体中的单倍型。结果表明：59571364位点的TT基因型和59912586位点的GG基因型在瘦尾中国美利奴与萨福克羊群体中属于优势基因型, 而两种基因型在脂尾(臀)阿勒泰羊与湖羊群体中比率均不足2%; 两位点在阿勒泰羊群体中的单倍型分析结果显示, CA单倍型为主单倍型, 比率高达55%, CA与TA两单倍型约占88.33%。以上结果提示, 绵羊X染色体59571364与59912586位点在脂尾(臀)与瘦尾绵羊群体中分布存在较大差异, 可作为理想的分子标记应用于高、低脂绵羊品种选育。     

7号染色体46765080位点SNP与绵羊尾臀性状相关性研究北大核心CSCD

绵羊尾(臀)脂性状是重要耐逆性状,其脂肪沉积的分子机制不清。旨在研究绵羊脂尾(臀)沉积脂肪的分子遗传机制,以最近文献报道的7号染色体一处SNP位点为候选分子标记,利用PCR-RFLP方法检测该位点在尾型极端差异的阿勒泰羊、哈萨克羊、湖羊、中国美利奴细毛羊以及萨福克羊群体中的多态性,并采用模型分析其与尾(臀)脂性状的相关性。结果表明,7号染色体46765080位点的G等位基因高频出现在表型分值较高的臀脂型阿勒泰羊群体中,A等位基因在长瘦尾绵羊品种中高频出现;等位基因频率G/A的比值与尾臀表型分值相关性模型表明G/A比值随着尾臀表型分值增加呈指数增长。以上结果表明,绵羊7号染色体46765080位点在尾(臀)脂与瘦尾绵羊群体中分布存在显著性差异,该SNP位点可作为一个理想的分子标记应用于高、低脂绵羊品种选育。     

新疆地区绵羊品种朊蛋白基因136、154和171位点的多态性

绵羊痒病是一种渐进性和致死性中枢神经系统疾病,绵羊朊蛋白基因(Prion protein gene,PRNP)多态性与痒病的易感或抗性有关,其中PRNP136位(V/A)、154位(H/R)和171位(H/Q/R)的基因多态性与该病发生最相关。为评价新疆地区主要绵羊品种对痒病的易感性,文章对新疆地区10个绵羊品种(阿勒泰、巴士拜、巴音布鲁克、多浪、和田、策勒黑、中国美利奴、德国肉用美利奴、特克赛尔和萨福克羊)共746只个体PRNP基因的136位(V/A)、154位(H/R)和171位(H/Q/R)的遗传多态性进行分析,检测到了ARQ、ARR、ARH、ARK、VRQ、AHR、AHQ、AHH8种等位基因,其中ARQ和ARR等位基因存在于所有品种中,且ARQ在所有品种中的基因频率最高。ARH存在于除萨福克和德国肉用美利奴羊外的8个绵羊品种中。仅在新疆地方品种阿勒泰、巴音布鲁克、巴士拜和多浪羊中检测到ARK等位基因。而VRQ、AHR、AHQ和AHH4种等位基因只在中国美利奴羊上存在,且频率极低。在10个品种中共检测到了ARQ/ARQ、ARQ/ARK、ARR/ARR、ARH/ARH、ARQ/ARR、ARH/ARQ、ARH/ARR、ARK/ARK、ARH/ARK、ARQ/VRQ、ARQ/AHQ、ARQ/AHR和ARH/AHH13种基因型,其中中度易感的ARQ/ARQ基因型频率最高,而抗性最强的ARR/ARR基因型仅存在于巴音布鲁克、策勒黑、中国美利奴、特克赛尔和德国肉用美利奴羊,且频率较低。文章首次在中国美利奴羊上发现了易感性很强的VRQ/ARQ基因型。上述结果提示新疆的主要绵羊品种对痒病的抗性较弱。     

FABP4基因在阿勒泰羊尾脂沉积与代谢模型中的表达变化规律

FABP4(Fatty acid binding protein 4)参与细胞内脂肪酸的转运和脂肪酸代谢,是当前研究动物脂肪沉积与代谢的热门候选基因。为了研究FABP4基因在绵羊尾脂沉积与代谢中的作用,文章采用生物信息学方法分析了FABP4氨基酸序列在各物种中的保守性;利用半定量RT-PCR方法检测了该基因在阿勒泰羊主要组织中的表达;采用饥饿法成功建立了模拟阿勒泰羊尾脂沉积与代谢的动物模型,利用qPCR和iTRAQ(isobaric tags for relative and absolute quantitation)技术同时验证FABP4基因mRNA和蛋白在阿勒泰羊非饥饿组与饥饿组尾脂中的表达变化。序列分析结果表明,绵羊FABP4氨基酸序列在物种间高度保守,提示FABP4基因可能由于其重要的生物功能而在进化中表现出其物种的保守性。组织表达谱结果显示,FABP4 mRNA在阿勒泰羊肠脂与尾脂中均高丰度表达,暗示FABP4基因可能在脂肪中行驶着重要的生理生化功能。qPCR与iTRAQ结果显示,FABP4 mRNA与蛋白在两种极端条件下尾脂中的表达差异均不显著(P>0.05),表明FABP4基因可能不是绵羊尾脂沉积与代谢两种极端差异表型的决定基因。以上研究结果为进一步研究FABP4基因在绵羊尾脂中的生物功能奠定了基础。     

金堂黑山半GOLA-DRB3基因多态性与体尺性状相关性

宁夏大家生命科学院杨易,徐金瑞,西南民族大学生命科学学院郑玉才,王杰四位动物遗传育种工作者采用TaqⅠ、PstⅠ和HaeⅢ三种限制性内切酶对111号金堂黑山羊GOLA-DKB3基因第二外显子的遗传多态性进行检测,其结果表明：金堂山羊GOLA—DRB3;第二外显子多态性极丰富,且为多碱基突变,除241位点外.     

中国美利奴羊H-FABP基因多态性及其与部分肉质性状相关性的研究

利用聚合酶链式反应-单链构象多态（PCR-SSCP）技术检测中国美利奴羊(Chinese Merino)心脏型脂肪酸结合蛋白基因（H-FABP）外显子2的单核苷酸多态性（SNPs）和遗传多态性,分析其与肌内脂肪（IMF）含量、肌纤维直径和肌纤维密度的相互关系,为该品种绵羊的分子标记辅助选择提供理论依据。结果显示,H-FABP基因外显子2有AA、AB和BB 3种基因型,AA型和BB型在778位均发生了C缺失,939位均发生了A→G转换,BB型还在789位发生了T→C转换,该突变导致所编码氨基酸发生了缬氨酸→丙氨酸的替换;BB型为IMF的优势基因型,与AB型相比差异显著(P<0.05),与AA型相比差异极显著（P<0.01);BB型对肌纤维直径存在负相关。结果提示,中国美利奴羊H-FABP基因外显子2具有多态性,该基因可能是中国美利奴羊肉质性状的主效基因,或者与控制这些性状的主效基因相连锁。     

脂蛋白脂酶基因多态性与2 型糖尿病的相关性研究

目的：探讨脂蛋白脂酶（lipoprotein lipase,LPL）基因PvuⅡ酶切多态性与2型糖尿病的相关性。方法：采用聚合酶链反应-限制性片段长度多态性（PGR-RFLP）方法,分析了156例样本LPL基因第6内含子PvuⅡ多态性（病例组98人。对照组58。其中40个2型糖尿病同胞对,病例组40人,对照组40人）。结果：病例组与对照组的基因型和基因频率均无显著性差异。结论：湖北汉族人群脂蛋白脂酶基因PvuⅡ酶切多态性与2型糖尿病无明显关联。     

中国美利奴羊H-FABP基因多态性及其与部分肉质性状的相关性

利用聚合酶链式反应-单链构象多态（PCR-SSCP）技术检测中国美利奴羊（Ovis aries var. Merino）心型脂肪酸结合蛋白基因（H-FABP）外显子2的单核苷酸多态性（SNPs）和遗传多态性,分析其与肌内脂肪（IMF）含量、肌纤维直径和肌纤维密度的相互关系,为该品种绵羊的分子标记辅助选择提供理论依据。结果显示,H-FABP基因外显子2有AA、AB和BB 3种基因型,AA型和BB型在778位均发生了C缺失,939位均发生了A→G转换,BB型还在789位发生了T→C转换,该突变导致所编码氨基酸发生了缬氨酸→丙氨酸的替换;BB型为IMF的优势基因型,与AB型相比差异显著（P<0.05）,与AA型相比差异极显著（P<0.01）;BB型对肌纤维直径存在负相关。结果提示,中国美利奴羊H-FABP基因外显子2具有多态性,该基因可能是中国美利奴羊肉质性状的主效基因,或者与控制肉质性状的主效基因相连锁。     

黄颡鱼MSTN基因多态性及其与生长性状的相关性分析

肌肉生长抑制素基因(Myostatin,MSTN)属于转化生长因子β家族,其主要功能是负向调控肌肉的生长发育。采用PCR-SSCP技术对黄颡鱼MSTN基因进行单核苷酸多态性检测和分型,并与其生长性状进行关联分析。结果表明,在第一内含子部分检测到1个缺失位点和2个突变位点(T1003del、G1022A和T1063G),基因型分别为:AA、AB、CC、CD和DD;在第三外显子部分检测到1个突变位点(T132C),基因型分别为:EE和EF。关联性分析表明,AA基因型个体的全长、体长、体高、体厚、头长和体重显著大于CD和DD基因型(P<0.05),AA基因型雌性个体的全长、体长、体高、体厚、头长、尾柄高、尾柄厚和体重也显著大于DD基因型(P<0.05)。由此推断,AA基因型是影响雌性黄颡鱼生长性状的有利基因型,DD基因型是影响雌性黄颡鱼生长性状的不利基因型,可以尝试利用这两个位点对雌性黄颡鱼进行标记辅助选育。     

Replication and refinement of a quantitative trait locus influencing milk protein percentage on ovine chromosome 3

A previous genome scan that was conducted in Spanish Churra sheep identified a significant quantitative trait locus (QTL) for milk protein percentage (PP) on chromosome 3 (OAR3), between markers KD103 and OARVH34. The aim of this study was to replicate these results and to refine the mapped position of this QTL. To accomplish this goal, we analysed 14 new half‐sib families of Spanish Churra sheep including 1661 ewes from 29 different flocks. These animals were genotyped for 21 microsatellite markers mapping to OAR3. In addition to a classical linkage analysis (LA), a combined linkage disequilibrium and linkage analysis (LDLA) was performed with the aim of enhancing the resolution of the QTL mapping. The LA that was performed in this sheep population identified the presence of a highly significant QTL for PP near marker KD103 (P_c < 0.001; P_exp < 0.001). The phenotypic variance that was owing to the QTL was 2.74%. Two segregating families for the target QTL were identified in this population with QTL effect estimates of 0.47 and 0.95 SD. The LDLA identified the same QTL as the previous analyses with a high level of statistical significance (P = 9.184 E‐11) and narrowed the confidence interval (CI) to a 13 cM region. These results confirm the segregation of the previously identified OAR3 QTL that influences PP in Spanish Churra sheep. Future research will aim to increase the marker density across the refined CI and to analyse the corresponding candidate genes to identify the allelic variant or variants that underlie this genetic effect.     

A single nucleotide polymorphism on chromosome 10 is highly predictive for the polled phenotype in Australian Merino sheep

The aim of this study was to fine map the genomic location of the Horns locus in the Australian Merino sheep population and to identify markers that can be used to predict the horn phenotype. A linkage disequilibrium analysis of horn data from Australian Merino sheep mapped the Horns locus to a small region on chromosome 10. A single nucleotide polymorphism in the region was found to be highly predictive for the polled phenotype in an experimental population of Merino sheep. This was owing to a dominance effect of one of the alleles when inherited maternally. It was suggested that a genetic test would provide a good predictor of the polled phenotype. Finally, an evaluation of industry data showed that the SNP is at very different frequencies in Poll Merino sheep that have been bred for polledness (based on phenotype alone) compared with the Merino sheep breed.     

Mapping of quantitative trait loci on porcine chromosome 4

A F₂ population derived from a cross between European Large White and Chinese Meishan pigs was established in order to study the genetic basis of breed differences for growth and fat traits. Chromosome 4 was chosen for initial study as previous work had revealed quantitative trait loci (QTLs) on this chromosome affected growth and fat traits in a Wild Boar × Large White cross. Individuals in the F₂ population were typed for nine markers spanning a region of approximately 124 c m . We found evidence for QTLs affecting growth between weaning and the end of test (additive effect: 43·4 g/day) and fat depth measured in the mid-back position (additive effect: 1·82 mm). There was no evidence of interactions between the QTLs and sex, grandparents or F₁ sires, suggesting that the detected QTLs were fixed for alternative alleles in the Meishan and Large White breeds. Comparison of locations suggests that these QTLs could be the same as those found in the Wild Boar × Large White cross.     

Genetic mapping of quantitative trait loci for aseasonal reproduction in sheep

The productivity and economic prosperity of sheep farming could benefit greatly from more effective methods of selection for year-round lambing. Identification of QTL for aseasonal reproduction in sheep could lead to more accurate selection and faster genetic improvement. One hundred and twenty microsatellite markers were genotyped on 159 backcross ewes from a Dorset × East Friesian crossbred pedigree. Interval mapping was undertaken to map the QTL underlying several traits describing aseasonal reproduction including the number of oestrous cycles, maximum level of progesterone prior to breeding, pregnancy status determined by progesterone level, pregnancy status determined by ultrasound, lambing status and number of lambs born. Seven chromosomes (1, 3, 12, 17, 19, 20 and 24) were identified to harbour putative QTL for one or more component traits used to describe aseasonal reproduction. Ovine chromosomes 12, 17, 19 and 24 harbour QTL significant at the 5% chromosome-wide level, chromosomes 3 and 20 harbour QTL that exceeded the threshold at the 1% chromosome-wide level, while the QTL identified on chromosome 1 exceeded the 1% experiment-wide significance level. These results are a first step towards understanding the genetic mechanism of this complex trait and show that variation in aseasonal reproduction is associated with multiple chromosomal regions.     

Purine nucleoside phosphorylase polymorphism in sheep erythrocytes

A polymorphism of purine nucleoside phosphorylase is described in sheep erythrocytes. Two isozymes were distinguished electrophoretically, one with high activity (NP-1) and one with low activity (NP-2). Breeding data suggest that the two isozymes are the product of two codominant alleles, NP ¹and NP ². The K _m 's for inosine did not differ between NP-1 and NP-2; however, NP-2 had a lower pH optimum and was relatively unstable when incubated at 48 C.Contribution No. 421-J, from the Department of Pathology, Kansas Agricultural Experiment Station, Manhattan, Kansas. Supported in part by USPHS Grants HL-70119 and HL 12072.     

Age-dependent quantitative trait loci affecting growth traits in Scottish Blackface sheep

To dissect age-dependent quantitative trait loci (QTL) associated with growth and to examine changes in QTL effects over time, the Gompertz growth model was fitted to longitudinal live weight data on 788 Scottish Blackface lambs from nine half-sib families. QTL were mapped for model parameters and weekly live weights and growth rates using microsatellite markers on chromosomes 1, 2, 3, 5, 14, 18, 20 and 21. QTL significance (using α = 0.05 chromosome-wide significance thresholds, unless otherwise stated) varied with age, and those for growth rate occurred earlier than equivalent QTL for live weight. A chromosome 20 QTL for growth rate was significant from 4 to 9 weeks (maximum significance at 6 weeks) and for maximum growth rate. For live weight, this QTL was significant from 8 to 16 weeks (maximum significance at 12 weeks). A nominally significant chromosome 14 QTL was detected for growth rates from birth to week 2 in the same families and location as an 8-week weight QTL. In addition, at the same position on chromosome 14, a QTL was significant for growth rate for 17–28 weeks (maximum significance at 24 weeks). A chromosome 3 QTL was significant for weights at early ages (birth to week 4) and a growth rate QTL on chromosome 18 was significant from 8 to 12 weeks. Fitting growth curves allowed the combination of information from multiple measurements into a few biologically meaningful variables, and the detection of growth QTL that were not observed from analyses of raw weight data. These QTL describe distinct parts of an animal's growth curve trajectory, possibly enabling manipulation of this trajectory.     

Diet-dependent obesity and hypercholesterolemia in the New Zealand obese mouse: identification of a quantitative trait locus for elevated serum cholesterol on the distal mouse chromosome 5

AIMS: New Zealand obese (NZO) mice exhibit a polygenic syndrome of obesity, insulin resistance, and hypercholesterolemia that resembles the human metabolic syndrome. This study was performed in order to locate genes responsible for elevated serum cholesterol and to compare their effects under a standard and high fat diet.METHODS: A backcross population of NZO with SJL mice (NZO x F1(SJL x NZO)) was generated. Mice were raised on a normal or high fat diet and were monitored for 22 weeks (body weight, serum cholesterol, and blood glucose). A genome-wide scan was performed by genotyping of approximately 200 polymorphic microsatellite markers by PCR and linkage analysis was performed with the MAPMAKER program.RESULTS: In the genome-wide scan, a single susceptibility locus for hypercholesterolemia (Chol1/NZO, maximum LOD score 14.5 in a combined population of 523 backcross mice) was identified on chromosome 5. Cholesterol levels were significantly elevated in both male and female homozygous carriers of the Chol1/NZO allele. The locus maps 40cM distal of the previously described obesity locus Nob1 in the vicinity of the marker D5Mit244 and in the vicinity of hypercholesterolemia QTL previously identified in the NZB, CAST, and C57BL/6J strains. Chol1/NZO was not associated with elevated body weight, serum insulin, or hyperglycemia. The high fat diet significantly increased serum cholesterol levels, but the fat content of the diet did not alter the absolute effect of Chol1/NZO.Conclusions: Chol1/NZO is a major susceptibility locus on the distal mouse chromosome 5, which produces gender-independent hypercholesterolemia in NZO mice. The effect of Chol1/NZO was independent of the dietary fat content and was not associated with the other traits of the metabolic syndrome. Thus, it is suggested that the responsible gene might be involved in cholesterol metabolism.