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为了筛选出高产娟姗牛的分子遗传标记,并应用这些分子标记进行高产娟姗牛的选育,笔者选用250条10碱基随机引物在由20头高产(305 d产奶量4000 kg)和20头低产(305 d产奶量3200 kg)娟姗牛所组成的2个DNA池间进行RAPD-PCR扩分子标记筛选。用筛选到的200条引物对所有个体进行PCR检测,得到了3个与奶牛产奶量性状相关的RAPD标记,并对它们进行了克隆测序以及生物信息学分析。试验结果表明,S11、S1110和S120 3个DNA片段与产奶量有关,其中S11与产奶量性状正相关关系,S1110和S120与产奶量性状呈负相关关系。 相似文献
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为能利用抗热应激SCAR标记应用于奶牛抗热应激新品种的选育,随机选取52头高产耐热荷斯坦奶牛和41头高产不耐热荷斯坦奶牛,利用RAPD技术对筛选出的耐热分子标记进行序列分析和SCAR转化,并以S441 SCAR标记对种公牛和核心群的母牛进行检测和分析。结果表明,随机引物S441和S463在高产耐热组中分别扩增了581 bp和680 bp的特异性片段,序列比较显示,前者与KCNN2基因的同源性达90%,后者与NW_001024067.1|BtUn_WGA9442_2同源性达99%;S441 SCAR标记在种牛的检出率为:母牛12.7%[27/213],公牛32.6%[15/46];数据统计分析表明:有无S441 SCAR标记的母牛产奶量差异不显著(P>0.05),但在热应激条件下产奶量下降率差异极显著(P<0.01)。研究初步确定S441标记作为荷斯坦奶牛抗热应激分子标记。 相似文献
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荷斯坦奶牛抗热应激SCAR标记的研究与应用 总被引：1，自引：0，他引：1
为能利用抗热应激SCAR标记应用于奶牛抗热应激新品种的选育,随机选取52头高产耐热荷斯坦奶牛和41头高产不耐热荷斯坦奶牛,利用RAPD技术对筛选出的耐热分子标记进行序列分析和SCAR转化,并以S441 SCAR标记对种公牛和核心群的母牛进行检测和分析.结果表明,随机引物S441和S463在高产耐热组中分别扩增了581 bp和680 bp的特异性片段,序列比较显示,前者与KCNN2基因的同源性达90%,后者与NW-001024067.1|BtUn-WGA9442_2同源性达99%;S441 SCAR标记在种牛的检出率为:母牛12.7%[27/213],公牛32.6%[15/46];数据统计分析表明:有无S441 SCAR标记的母牛产奶量差异不显著(P>0.05),但在热应激条件下产奶量下降率差异极显著(P<0.01).研究初步确定S441标记作为荷斯坦奶牛抗热应激分子标记. 相似文献
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选取高产荷斯坦牛10头，优化奶牛RAPD反应最佳体系，并在154条随机引物中筛选出稳定性好、带型清晰的引物64条。结合红细胞钾测定结果和生产性能测定，通过RAPD分析并筛选出3个与奶牛耐热性状相关的分子标记，以期应用这些分子标记进行高产耐热荷斯坦奶牛的选育工作。 相似文献
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选取10头高产荷斯坦牛为实验材料,优化奶牛RAPD反应最佳体系,并在154条随机引物中筛选出稳定性好、带型清晰的引物64条。结合红细胞钾测定结果和生产性能测定,通过RAPD分析并筛选出3个与奶牛耐热性状相关的分子标记,以期应用这些分子标记进行高产耐热荷斯坦奶牛的选育。 相似文献
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家蚕雌特异分子标记筛选、克隆及其序列分析 总被引：1，自引：0，他引：1
为了建立家蚕W染色体的特异性分子标记 ,采用RAPD PCR技术对同一品种家蚕雌雄个体基因组DNA进行分子标记筛选。通过 4 5 0种随机引物筛选到 1条雌特异性的DNA片段OPZ0 7 35 5bp ,将该雌特异片段克隆到载体pGEM T上 ,并进行测序和同源性分析 ,结果表明 :该雌特异DNA片段的序列长为 35 5bp ,推测其可能来自于家蚕的W染色体。 相似文献
10.
奶牛外貌评分与产奶量相关性研究 总被引：1，自引：0，他引：1
某奶牛场随机取30头奶牛,测定其305d产奶量及10个体型外貌评分性状,计算产奶量与体型性状之间的相关系数,求得产奶量的各自通径系数,研究奶牛10个体型性状与生产效率性状之间的关系。结果表明,整体评分与305d产奶量之间存在弱正相关（P〉0．05）;10个评分性状中,后乳房高度和宽度与305d产奶量呈极显著相关（P〈0．01）。乳房中隔和乳房深度与305d产奶量呈显著相关（P〈0．05）。整体外貌评分、后乳房宽度、乳房中隔越高则产奶量也越高,后乳房高度和乳房中隔是对产奶量起主要作用的性状。 相似文献
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An independent confirmation of a quantitative trait locus for milk yield and composition traits on bovine chromosome 26 总被引：1，自引：0，他引：1
Z. Jiang S. De M.D. Garcia K.B. Griffin X.-L. Wu Q. Xiao J.J. Michal B.S. Sharma & G.B. Jansen 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2005,122(4):281-284
Several reports have demonstrated that bovine chromosome 26 (BTA26) harbours significant or suggestive quantitative trait loci (QTL) for milk production and composition traits in dairy cattle. Our previous study showed that a C/T substitution in the bovine TCF7L2 gene on BTA26 was significantly linked to QTL for protein yield (PY) in a Canadian dairy cattle population. Actually, this polymorphism was one of the markers derived from a genome‐wide screening of QTL for milk PY using an amplified fragment length polymorphism technique combined with a DNA pooling strategy. In the present study, 990 Holstein bulls with complete genotype and phenotype data from 14 sire families were analysed to confirm, if the QTL effects exist in other populations. Statistical analysis revealed that this marker was significantly associated with PY, protein percentage, milk yield and fat yield (FY) (p < 0.001) in the US Holstein population. These results indicate that this QTL region has a pleiotrophic effect on different milk traits and is portable in different populations. 相似文献
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BCA Alves VFM Hossepian de Lima CA Moreira‐Filho 《Reproduction in domestic animals》2010,45(6):1047-1051
Sex pre‐selection of bovine offsprings has commercial relevance for cattle breeders and several methods have been used for embryo sex determination. Polymerase chain reaction (PCR) has proven to be a reliable procedure for accomplishing embryo sexing. To date, most of the PCR‐specific primers are derived from the few single‐copy Y‐chromosome‐specific gene sequences already identified in bovines. Their detection demands higher amounts of embryonic genomic material or a nested amplification reaction. In order to circumvent this, limitation we searched for new male‐specific sequences potentially useful in embryo sexing using random amplified polymorphic DNA (RAPD) analysis. Random amplified polymorphic DNA (RAPD) assay reproducibility problems can be overcome by its conversion into Sequence Characterized Amplified Region (SCAR) markers. In this work, we describe the identification of two bovine male‐specific markers (OPC16323 and OPF101168) by means of RAPD. These markers were successfully converted into SCARs (OPC16726 and OPF10984) using two pairs of specific primers.Furthermore, inverse PCR (iPCR) methodology was successfully applied to elongate OPC16323 marker in 159% (from 323 to 837 bp). Both markers are shown to be highly conserved (similarity ≥95%) among bovine zebu and taurine cattle; OPC16323 is also highly similar to a bubaline Y‐chromosome‐specific sequence. The primers derived from the two Y‐chromosome‐specific conserved sequences described in this article showed 100% accuracy when used for identifying male and female bovine genomic DNA, thereby proving their potential usefulness for bovine embryo sexing. 相似文献
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Zhang CL Chen H Wang YH Zhang RF Lan XY Lei CZ Zhang L Zhang AL Hu SR 《Research in veterinary science》2008,85(2):265-268
Serotonin receptor 1B (HTR1B) is one of the 14 different identified serotonin receptors which are involved in the regulation of behaviors such as sleep, fear, aggression, mood, and feeding. The aims of this study were to characterize polymorphisms in the 5' coding and the 3' flanking regions of the bovine HTR1B gene among Chinese Holsteins and to identify the association of HTR1B polymorphisms with milk production performance. SSCP was used to examine the polymorphisms at four loci and the fragments with different SSCP patterns were sequenced. A total of three single nucleotide polymorphisms (SNPs) were detected, among which the 205G>T mutation was found to cause a predicted amino acid change: from Ala to Ser at position 69. The polymorphism of G205T was identified to be associated with milk yield trait. Furthermore, the H1-C genotype was found to be associated with a significant increase in milk yield of 489kg vs. the H1-A genotype (P<0.05). 相似文献
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中国黄牛PrPc成熟蛋白基因的克隆和序列分析 总被引：3，自引：0，他引：3
根据已发表的PrP^c蛋白基因序列，设计合成一对引物，并在2个引物的两端分别加入Bam HI和Nde Ⅰ酶切位点。以从中国黄牛肝脏中提取的基因组DNA为模板，经PCR扩增出一约640bp的目的基因片段，将此基因克隆于pET11a载体，构建了PrP^c蛋白基因重组质粒b-pET11a-PrP^c，转化DH－5α并进行序列测定。同源性分析表明：中国黄牛PrP^c成熟蛋白基因与目前国外发表牛的成熟蛋白PrP^c基因相比，有较大差异，发现了一个新的NdeⅠ酶切位点；推导的氨基酸序列有4个氨基酸差异，缺少一个8氨基酸重复区。 相似文献
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荷斯坦奶牛STAT5A基因的SNPs检测及其与产奶性状的关联分析 总被引：4，自引：0，他引：4
根据GenBank公布的牛STAT5A基因的2个SNPs（AJ237937和AF079568）的引物序列,采用PCR-SS-CP方法对758头中国荷斯坦奶牛进行了STAT5A基因多态性检测,并将其与5个产奶性状进行了关联分析。在SNP1的9501碱基处发现A→G突变;SNP2中发现2个连锁点突变,即12 440位T→C和12 550位的CCT插入/缺失;方差分析结果表明：SNP1对乳蛋白率有显著影响（P〈0.05）;SNP2对产奶量有极显著影响（P〈0.01）,对乳脂量、乳蛋白量有显著影响（P〈0.05）;单倍型组合对产奶量和乳蛋白量有极显著影响（P〈0.01）,对乳脂量有显著影响（P〈0.05）。多重比较分析表明SNP2的AB基因型的产奶量、乳脂量和乳蛋白量的最小二乘均数极显著（P〈0.01）或显著（P〈0.05）高于基因型AA;而单倍型组合H1H2的产奶量、乳脂量和乳蛋白量的最小二乘均数显著（P〈0.05）或极显著（P〈0.01）高于其它4种单倍型组合;H3H4对产奶量和乳蛋白量的效应极显著高于其它4种单倍型组合（P〈0.01）。结果提示：STAT5A基因可以作为奶牛产奶性状的候选分子遗传标记。 相似文献
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以51头西门塔尔和鲁西黄牛的杂交后代作为试验动物,扩增组织蛋白酶D(cathepsin D,CTSD)基因部分外显子,通过DNA测序方法检测到该基因第5、6、8外显子共4个SNPs(T581C、T599C、A861T、A1128G)的遗传变异。SAS软件采用最小二乘法拟合线性模型,将几个位点的不同基因型与牛肉质、胴体性状进行关联分析。结果表明,位于CTSD基因外显子5的T581C突变与大理石纹性状呈显著相关(P<0.05),BB基因型显著优于AA和AB基因型,这为T581C作为肉牛大理石纹的分子标记提供了一定的理论依据。 相似文献
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E. Bagnicka N. Strza&#;kowska K. Flisikowski T. Szreder A. Jó&#;wik B. Prusak J. Krzy&#;ewski & L. Zwierzchowski 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2007,124(3):150-156
The aim of this study was to find a polymorphism of the bovine β4‐defensin gene and search for its association with milk yield and composition and with the somatic cell count in milk. The data were from the years 1999 to 2004 on 212 Holstein‐Friesian (HF) dairy cows, descended from 70 sires. Based on the sequence of the bovine β4‐defensin gene (GenBank no. AF008307 ) the primers were designed for the amplification of the 924‐bp or 393‐bp long fragments. The 924‐bp long fragment was sequenced and the sequence was compared with that available in the GenBank. Ten putative nucleotide sequence polymorphisms were found in the intron of the bovine β4‐defensin gene. One of them, a C→T transition at position 2239, that creates a new NlaIII (Hin1II) restriction site, was genotyped with polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) in a cohort of 212 HF cows. The CC genotype was the most common (72%). The heterozygous CT genotype was found in 26% of the genotyped cows and four cows (2%) were TT homozygotes. In order to determine the relationship between the polymorphism of the β4‐defensin gene and milk production traits a multi‐trait repeatability test‐day animal model was used. The Derivative‐free Multivariate analysis program was used for computation. The differences between estimates for genotypes were checked using Student's t‐test. The model included the animal genotype, year‐season of calving and parity as fixed effects and the animal additive genetic effect and permanent environmental effect of individual cows as well as dates of the tests as random effects. Significant associations were found between the RFLP‐NlaIII and milk fat, protein and lactose contents. Also, a significant effect was shown of the defensin genotype on the somatic cell count in the milk. 相似文献
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用40条10碱基随机引物,对普通小麦“中国春”、“中国春”-杀配子染色体2C二体附加系、柱穗山羊草进行RAPD分析,筛选到1个杀配子染色体2C特异引物OPF03。从“中国春”-杀配子染色体2C二体附加系中克隆了该DNA片段。测序结果表明,该片段长1 496 bp,与大麦1个cDNA的同源性为92%。根据OPF031496的序列设计了1对SCAR引物2C-F586、2C-R586,对普通小麦“中国春”、“中国春”-杀配子染色体2C二体附加系、柱穗山羊草、二倍体长穗偃麦草、“中国春”-长穗偃麦草7E二体附加系等材料进行了SCAR分析,在“中国春”-杀配子染色体2C二体附加系、柱穗山羊草中扩增出了586 bp的片段,而在普通小麦“中国春”、二倍体长穗偃麦草、“中国春”-长穗偃麦草7E二体附加系中没有该产物,初步证明此标记可以用于杀配子染色体2C的检测。进一步用该对SCAR引物对“中国春”-杀配子染色体2C二体附加系与“中国春”-长穗偃麦草7E二体附加系的杂种F1代、F2代进行了分析,结果在全部F1代以及部分F2代材料扩增出了目的片段,进一步证明了此SCAR标记可以用来检测小麦背景下的杀配子染色体2C,为小麦背景中杀配子染色体2C的快速跟踪检测提供了新途径。 相似文献
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作者所在团队前期通过奶牛乳腺上皮组织转录组测序及荷斯坦公牛全基因组重测序研究发现RPL23A和ACACB基因是奶牛乳蛋白和乳脂性状的候选功能基因,本研究旨在探究这两个基因是否对奶牛产奶性状具有显著遗传效应。以北京地区7个牧场的1059头中国荷斯坦母牛为试验群体,采集尾根静脉血并提取基因组DNA,通过飞行时间质谱方法检测SNP位点基因型,利用SAS9.4软件的MIXED过程进行关联分析。结果表明,RPL23A基因的SNP位点g.20146771C>T与第1泌乳期5个产奶性状达到显著或极显著关联(P=0.0001~0.0416),其优势等位基因为T;ACACB基因的g.63878254T>C位点与第1泌乳期产奶量、乳脂量和乳蛋白量呈极显著关联(P<0.01),其优势等位基因为C;g.63962768G>A位点与第1泌乳期产奶量、乳脂量、乳脂率和乳蛋白率关联显著或极显著(P=0.0001~0.0391),其优势等位基因为A。综上,RPL23A基因主要影响中国荷斯坦牛产奶量和乳蛋白,ACACB基因对产奶量和乳脂具有显著遗传效应,3个SNP位点可考虑作为遗传标记用于标记辅助选择培育奶牛高乳蛋白乳脂新品系和选育提高。 相似文献
20.
Liefers SC Veerkamp RF Te Pas MF Chilliard Y Van der Lende T 《Domestic animal endocrinology》2005,29(1):227-238
In dairy cattle, the increase in milk yield has been accompanied by a more negative energy balance (EB) during early lactation and a decrease in fertility. As the hormone leptin is involved in regulation of nutritional status and reproductive function this hormone is an interesting protein to investigate during the periparturient period in dairy cattle. This study was performed to get insight into the function of leptin during the periparturient period and to perform an association study between polymorphisms in the bovine leptin gene and leptin receptor gene and fertility as well as production traits. Leptin concentrations in the periparturient cow undergo remarkable changes; leptin concentrations were high during late pregnancy and declined to a nadir at parturition. Genetic analysis of the leptin gene indicated that a combination of three polymorphisms located at a 135 bp region of the leptin promoter explained most of the variance in prepartum leptin concentrations. The two extreme genotype combinations could be used to investigate the function of leptin concentrations in pregnant cows. A polymorphism located on intron 2 of the leptin gene explained a significant part of the variation in milk yield. On the promoter region of the leptin gene an SNP was detected that was associated with first postpartum luteal activity (FPLA). This SNP could be a candidate marker for fertility in dairy cows. Another SNP on the leptin promoter was associated with energy balance and dry matter intake (DMI) where a higher dry matter intake occurred together with a higher energy balance. Two genotype combinations of the aforementioned three associated SNPs were defined which had a good milk yield together with a good energy balance and fertility. Calculations of an economical value per trait have to validate if one of these genotype combinations would be a possible candidate to be used in selection. 相似文献