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为了构建北京奶牛中心荷斯坦种公牛个体及亲子鉴定DNA数据库,本研究从联合国粮农组织(FAO)和国际动物遗传学会(ISAG)推荐的具有高度多态性的微卫星标记中选取11个常染色体微卫星标记(BM1824、BM2113、ETH3、ETH10、ETH225、INRA023、SPS115、TGLA53、TGLA122、TGLA126和TGLA227),采用荧光引物PCR和ABI3730XL遗传分析仪,对229头荷斯坦种公牛个体基因型进行了检测,并探讨其用于牛个体识别和亲子鉴定的可行性。研究结果表明,11个微卫星标记平均杂合度为0.733,平均多态信息含量为0.695,其中TGLA122标记平均多态信息含量最高为0.866,SPS115标记最低为0.506,均属于高多态性标记。11个标记累积个体识别能力为99.99%,累积非父排除率达到99.99%,随机个体一致性概率为1.59×10-17。研究结果表明上述11个微卫星标记进行个体识别和亲子鉴定的效力与可靠性均很高,同时初步建立了北京奶牛中心荷斯坦种公牛DNA数据库。 相似文献
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利用微卫星和SNP标记信息进行奶牛亲子鉴定的模拟研究 总被引：2，自引：0，他引：2
本研究旨在比较微卫星标记和单核苷酸多态(SNP)标记进行奶牛亲子鉴定的效率。研究中随机模拟了10 000对含微卫星或SNP标记信息的非亲子关系的个体,用排除法进行亲子鉴定,该过程重复100次。结果表明,标记的多态性对于排除概率的影响最大。达到同样的排除概率,所需低多态标记数量远高于高多态标记。当真实母亲基因型已知时,个体父亲的推断效率较高。达到同样的排除概率,无论标记多态性高或低,3种鉴定类型所需SNP标记数量均大于微卫星标记。当SNP和微卫星标记的平均杂合度为0.484和0.875时,需要的SNP标记数是微卫星标记的5~6倍;当SNP和微卫星标记的平均杂合度为0.363和0.566时,需要的SNP标记数约是微卫星标记的2倍。在2种标记中,达到同样排除概率(大于0.99),采用至少2个矛盾推断原则推断所需标记数为单个矛盾推断原则的1.45倍。结果表明,由于SNP标记具有误判率低、重复性高和易于高通量自动化检测等特点,随着检测成本降低,SNP标记将来完全有可能取代微卫星标记用于奶牛群体的亲子鉴定。 相似文献
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为了构建基于微卫星标记的中国荷斯坦种公牛亲子鉴定及个体识别技术平台,本研究选取国际动物遗传学会（ISAG）推荐的12个微卫星标记（BM1824,BM2113,SPS115,ETH10,TGLA122,ETH225,INRA023,TGLA126,TGLA227,BM1818,ETH003和TGLA53）,采用荧光标记毛细管电泳方法,对571头荷斯坦种公牛进行基因型检测。统计结果显示,标记的平均多态信息含量为0．700,平均杂合度为0．735,属于高多态性标记;12个标记的累积排除概率大于0．996,累积一致性概率为4．9×10^-13,将其运用于亲子鉴定和个体识别具有很强的检测效率。此外．通过分析每个标记的检测信号峰图特征,将检测结果与商业化试剂盒进行比较以及利用ISAG参考DNA样品进行等位基因统一命名等手段,建立了标准化的微卫星等位基因分型方法,并初步建立起中国荷斯坦种公牛遗传检测信息库。 相似文献
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肉牛杂交亲本遗传距离与杂种优势的相关性研究 总被引：2，自引：0，他引：2
利用11个微卫星标记研究川南山地黄牛、海福特牛、黑安格斯牛、红安格斯牛、利木赞牛、西门塔尔牛、德国黄牛的遗传多样性,分析肉牛亲本间微卫星标记遗传距离与体尺、体重杂种优势间的相关性。结果表明:11个微卫星基因座平均有效等位基因数为4.5422,7个牛种的多态信息含量在0.3434~0.4207,平均杂合度为0.6868~0.8413,7个牛种均具有丰富的多态性。亲本间遗传距离与体尺、体重杂种优势间相关系数偏小(-0.003~0.304),均未达到显著水平,用微卫星标记遗传距离预测肉牛体尺、体重杂种优势还有待进一步探讨。 相似文献
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《中国畜牧杂志》2018,(12)
本研究旨在为猪亲子鉴定提供一种快速、准确的鉴定方法。先用非变性聚酰胺凝胶电泳方法对猪的55个微卫星位点进行筛选,再以高分辨率熔解曲线(HRM)技术对筛选出的微卫星位点进行多态性检测,再对其中一微卫星位点进行克隆测序验证,处理得到的数据,进行猪亲子鉴定研究。结果表明:利用非变性聚酰胺凝胶电泳实验筛选出12个适于HRM检测的微卫星位点,HRM技术对12个微卫星多态性进行检测,共检测出50个等位基因,平均值为4.167个,有效等位基因数平均值为2.675 0,群体遗传杂合度平均值为0.595 0,平均多态信息含量为0.540 0;对SW72位点克隆测序的结果与HRM技术检测结果一致;12个微卫星组合在双亲未知、只知单亲和双亲已知的情况下计算的累积排除率各为0.940 1、0.991 5和0.999 9;利用建立的亲子鉴定体系对样本中8个家系进行检测,结果累积排父率均大于0.999 9。HRM技术对12个微卫星位点组合进行检测,能够快速、准确地对猪进行亲子鉴定。 相似文献
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《畜牧兽医学报》2017,(4)
旨在利用较少的经过优化的遗传标记组合达到快速高效的亲缘鉴定,最终可有效应用于奶牛群体亲子鉴定与系谱构建。本研究从国际动物遗传学会(ISAG)数据库中初步选择了18个多态性较高的微卫星位点,经PCR优化验证获得可高效扩增的8个位点,并检测其在300头中国荷斯坦牛群体中的多态性分布。这些微卫星位点的亲子鉴定效力检测结果表明,8个标记平均等位基因数为14.63,平均多态性信息含量(PIC)为0.747,群体平均观察杂合度(Ho)为0.718,平均期望杂合度(He)为0.773。3种不同情形下进行亲子鉴定时8个位点的结合排除概率(CPE)均≥0.990。8对父子牛亲权鉴定结果表明,利用该8个标记可以识别并剔除系谱中的错误记录。结合排除概率的分析结果表明,当微卫星位点数增加至4个(TGLA227、TGLA122、BMC1207、BM103)时,3种不同情形下的CPE均≥0.949,可以满足个别案例牛亲子鉴定的要求。群体分子系谱构建的结果表明,当微卫星位点数大于或等于6个时,群体内近交系数和总群体近交系数均明显升高。因此,在生产实践中推荐使用TGLA227、TGLA122、BMC1207、BM103、INRA037、INRA134位点进行小规模群体(200)的分子系谱构建或亲权鉴定。 相似文献
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4个肉牛品种微卫星多态性分析 总被引：1，自引：1，他引：0
本研究探讨了引进牛种（西门塔尔牛、利木赞牛）和山东地方黄牛（鲁西黄牛、利鲁牛）在20个微卫星位点上的多态性。试验从不同地区的种公牛站采集鲁西黄牛、利鲁牛、利木赞牛和西门塔尔牛的血液或精液样本,分别为56、27、31和30个,共计144个样本,血液样本采用Lab-Aid基因组DNA分离试剂盒提取基因组DNA,精液样本采用高盐法提取基因组DNA。利用20个微卫星标记,采用荧光标记毛细管电泳方法,对4个肉牛品种的DNA多态性进行分析,主要研究了每个群体的遗传变异指标（等位基因数、多态信息含量和杂合度）及群体间的遗传关系（F-统计量和基因流）。结果表明,肉牛基因组DNA条带整齐,无拖尾现象,质量较好,可用于本试验。20个微卫星座位中共检测到211个等位基因,平均等位基因数为10.6个。群体的平均观测杂合度在0.6147~0.7176之间,平均期望杂合度在0.6735~0.7862之间。在所有群体中各位点的多态信息含量在0.4853~0.8714之间,平均为0.7284,但在各个群体内的差异较大。近交系数及基因流分析结果表明,4个肉牛品种近交系数较低,群体间平均近交系数为0.085,每个微卫星位点的基因流均>1。总体来看,所选用的20个微卫星座位多态信息含量高,可作为有效遗传标记用于肉牛品种间遗传多样性分析。 相似文献
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4个肉牛品种微卫星多态性分析 总被引：1，自引：1，他引：0
本研究探讨了引进牛种(西门塔尔牛、利木赞牛)和山东地方黄牛(鲁西黄牛、利鲁牛)在20个微卫星位点上的多态性。试验从不同地区的种公牛站采集鲁西黄牛、利鲁牛、利木赞牛和西门塔尔牛的血液或精液样本,分别为56、27、31和30个,共计144个样本,血液样本采用Lab-Aid基因组DNA分离试剂盒提取基因组DNA,精液样本采用高盐法提取基因组DNA。利用20个微卫星标记,采用荧光标记毛细管电泳方法,对4个肉牛品种的DNA多态性进行分析,主要研究了每个群体的遗传变异指标(等位基因数、多态信息含量和杂合度)及群体间的遗传关系(F-统计量和基因流)。结果表明,肉牛基因组DNA条带整齐,无拖尾现象,质量较好,可用于本试验。20个微卫星座位中共检测到211个等位基因,平均等位基因数为10.6个。群体的平均观测杂合度在0.6147~0.7176之间,平均期望杂合度在0.6735~0.7862之间。在所有群体中各位点的多态信息含量在0.4853~0.8714之间,平均为0.7284,但在各个群体内的差异较大。近交系数及基因流分析结果表明,4个肉牛品种近交系数较低,群体间平均近交系数为0.085,每个微卫星位点的基因流均1。总体来看,所选用的20个微卫星座位多态信息含量高,可作为有效遗传标记用于肉牛品种间遗传多样性分析。 相似文献
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OBJECTIVE: To evaluate a rapid polymerase chain reaction (PCR) fingerprinting technique for discriminating among Pasteurella multocida isolates from laboratory rabbits. SAMPLE POPULATION: 33 P multocida isolates from rabbits with clinical pasteurellosis. PROCEDURE: PCR assays were conducted with 2 minisatellites (core sequence and modified core sequence of phage M13) and 2 microsatellites ([GTG]5 and [GACA]4). Each bacterium was assigned to a PCR type for each of the primers used. Boiled bacterial extracts and purified genomic DNA were compared by use of PCR assays for phage M13 and (GACA)4. Plasmids were isolated from each bacterium, and their influence on PCR fingerprint was determined, using boiled extracts as a DNA source. RESULTS: M13 core sequence and M13 modified core sequence yielded 5 and 8 PCR types, respectively. The microsatellites (GTG)5 and (GACA)4 yielded 4 and 9 PCR fingerprint types, respectively. Fingerprint patterns obtained by use of isolated DNA differed from those obtained by use of boiled extracts, although discrimination among P multocida isolates was similar. The presence or absence of plasmids did not affect PCR fingerprints. CONCLUSION: Single primer PCR fingerprinting with minisatellite and microsatellite primers is an efficient and reproducible method for the discrimination of P multocida isolates from rabbits and can be performed directly, using boiled bacterial extracts as a source of template, although more bands were obtained from pure genomic DNA. 相似文献
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选取27个微卫星位点,应用PCR技术对贵州白香猪的2个品系进行遗传质量监测。结果在所选的27个微卫星位点中,共有24个位点获得稳定的扩增结果,其中SW911位点与S0026两个位点在群体内表现为单态纯合。2个品系有效等位基因、平均多态信息含量、平均杂合度、基因纯合率和平均近交系数分别为1.8310、1.7951、0.3436、0.3249、0.4329、0.4188、52.63%、58.55%、0.5377、0.5605。结果表明微卫星标记可用于检测贵州白香猪的遗传质量,也可用于分析封闭群的遗传多态性。是一种有潜力的试验动物遗传学检测标记。 相似文献
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Junghwa An Sung-Kyoung Choi Julie Sommer Edward Louis Jr Rick Brenneman Barbora Zemanov�� Petra H��jkov�� Grimm Park Mi-Sook Min Kyung-Seok Kim Hang Lee 《Journal of veterinary science (Suw?n-si, Korea)》2010,11(4):351-353
In order to screen microsatellites for conservation genetics studies of the species, a total of 23 microsatellite loci from Korean goral (Naemorhedus caudatus), including 15 previously developed loci and 8 new loci in this study, were tested. Eleven microsatellites were screened and subjected to cross-species amplification using a test panel of four Caprinae species, Japanese serows (Capricornis crispus), Chinese gorals (Naemorhedus goral), Northern chamois (Rupicapra rupicapra) and domestic goats (Capra hircus). In addition, all eleven microsatellites (SY3A, SY12A, SY12B, SY48, SY58, SY71, SY76, SY84, SY84B, SY112, and SY129) satisfied the criteria to be a core set of microsatellites. This core set of microsatellites and cross-species amplification of Korean goral microsatellites were found to be helpful for high-resolution studies for conservation and management of Korean goral and other endangered Caprinae species. 相似文献
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封闭群 Wistar大鼠的微卫星 DNA遗传监测分析 总被引：1，自引：0，他引：1
随机选取9只SPF级Wistar大鼠,其中4只雌性和5只雄性.在大鼠染色体上筛选了30个基因座位点,利用PCR扩增技术对封闭群Wistar大鼠群体进行了微卫星DNA多态性分析.结果有30对引物经扩增后均有图带产生,没有无效基因存在,图带均为双带.不同个体间的相似系数主要分布在0.3～0.8之间,在0.5～0.8之间占总数的94.4%,最高值为0.733.9号个体与其余个体间相似系数相对低于其他个体间的相似系数.筛选出了19个微卫星位点具有显著多态性,为建立一种对Wistar大鼠进行准确可靠、快速简便的遗传监测方法提供了依据. 相似文献
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Hiroki HIRAYAMA Akira FUJIKAWA Soichi KAGEYAMA Satoru MORIYASU Ken SAWAI Sadao ONOE Akira MINAMIHASHI 《Animal Science Journal》2008,79(5):554-560
This study examined genetic diagnosis using whole genome amplification (WGA) in bovine embryos. The first experiment was conducted to compare the WGA efficiency of primer extension preamplification-PCR (PEP-PCR) and multiple displacement amplification (MDA), and to optimize the DNA extraction method. The sensitivity of SRY -specific PCR from MDA products increased when DNA of fibroblasts was extracted by a NaOH treatment instead of the conventional method (heat treatment). The detectability of SRY from PEP-PCR products was lower than that in MDA regardless of the DNA extraction method (proteinase K or NaOH treatment). Sexing and genotyping were performed using MDA products from embryo biopsy. The accuracy of PCR-based and LAMP-based sexing was 100% regardless of the amounts of biopsy. Genotyping of CL16 , BND3 , SCD and F11 in MDA products from 10 to 20% of trophectoderm was successful 97, 97, 95 and 95% of the time, respectively, but reduced biopsy amount (<10% of trophectoderm) decreased the accuracies (33–83%). Microsatellite markers were analyzed using MDA products from 10 to 20% of trophectoderm. In eight out of 16 microsatellites, genotypes were not contradictory among the dam, sire and embryos. In the other eight microsatellites, the inconsistency rates were 17–83%. These results indicate that MDA is useful for multiple genetic diagnoses in bovine embryos. 相似文献
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Gomez-Raya L Priest K Rauw WM Okomo-Adhiambo M Thain D Bruce B Rink A Torell R Grellman L Narayanan R Beattie CW 《Journal of animal science》2008,86(1):17-24
The feasibility and economic value of DNA paternity identification were investigated and illustrated using Nevada beef cattle operations. A panel of 15 microsatellites was genotyped in 2,196 animals from 8 ranches with a total of 31,571 genotypes. Probabilities of exclusion for each marker within ranch and across ranches were computed. Joint probabilities of exclusion for the 15 microsatellites were also determined, resulting in values over 0.99 for any individual ranch and across ranches. Dropping 1 or 2 microsatellites with the lowest probabilities of exclusion resulted in joint probabilities greater than 0.99 and with marginal reduction compared with the probabilities with 15 microsatellites. Formulas for benefit-cost analysis for a DNA paternity identification program in beef cattle were derived. Genotyping 15 microsatellites with 20 calves per sire resulted in benefits of $1.71 and $2.44 per dollar invested at bull culling rates of 0.20 and 0.30, respectively. The breakpoints for the program to be profitable occurred when the ratio of the price of 1 kg of calf liveweight over the cost of genotyping 1 microsatellite was greater than 1.1 for a bull culling rate of 0.30. Benefit-cost analysis was also derived under incomplete DNA paternity identification using a lower number of DNA markers than necessary to achieve joint probabilities of exclusion of 0.99. Approximately a 20% increase in the benefit-cost ratio was achieved using 10 vs. 12 microsatellites with incomplete paternity identification. The greater the number of bulls in the operation, the lower the benefit-cost ratio of the paternity testing program. Low probabilities of exclusion and a high number of bulls in the beef operation reduced the benefit-cost ratio dramatically. The DNA paternity identification programs are feasible and may be profitable for free-range beef cattle operations. 相似文献
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Construction of a canine bacterial artificial chromosome library for screening with PCR 总被引：1，自引：0，他引：1
C. Schelling J. Schläpfer A. Billault K. Guziewicz A. Gmür I. Katmann B. Pineroli B. Colomb O. Rickli C. Wittwer A. Piasecka G. Dolf 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2002,119(6):400-401
We have constructed a canine bacterial artificial chromosome library amenable to PCR screening. The library consists of 96 768 clones and was initially screened with 112 microsatellites representing all canine chromosomes. For 87 primer sets (77%) one to seven positive superpools were identified. The library will be expanded by adding additional superpools in order to increase the genome coverage. Interested researchers can access the library following the rules published at http://www.dogmap.ch . 相似文献