【作者】 贾名威；

【导师】 杨利国；

【作者基本信息】 南京农业大学 ， 动物遗传育种与繁殖， 2003， 硕士

【摘要】 随着分子生物学的发展，遗传标记的概念发展为在核酸分子水平上具有相对差异的等位区域。STR基因座遵循孟德尔遗传规律，在基因组中分布广泛，具有PCR扩增，电泳分型等优点，在生物学各领域中得到广泛应用。在亲权鉴定、个体识别应用中，由于STR系统的使用，使检测进入了一个新的阶段，检测水平有了空前的提高。本研究的目的在于：①分析奶牛和肉牛6个STR基因座遗传多态性；⑦用微卫星标记技术建立准确快捷的亲权鉴定方法。 1 奶牛和肉牛6个STR基因座遗传多态性研究 试验所用样本采自两个品种，共161个样本。其中9份中国荷斯坦奶牛种公牛精液样本，23个家系中49份中国荷斯坦奶牛（母）血液样本；103份西门塔尔等肉牛品种种公牛精液样本。分别提取DNA，应用6对微卫星引物对奶牛和肉牛进行遗传多态性研究。利用PCR技术和复合电泳银染技术检测奶牛和肉牛BM2113、BM1862、BMc701、BM2934、TGLA122、BM720等6个STR基因座的多态性分布，并计算该6个基因座的基因频率（Pi）、个体鉴别力（DP）、杂合度（H）、多态信息含量（PIC）、和非父排除概率（PE）。结果显示：6个STR基因座的基因型分布符合Hardy-Weinberg平衡，奶牛中6个STR基因座中BM2113基因座的DP、H和PIC最高，TGLA122基因座的PE最高。6个STR基因座的累积个体鉴别力(C_DP为0.99997，累积非父排除能力(C_PE)为0.98827。肉牛中6个STR基因座中BM1862基因座的DP、H、PIC、PE都是最高，6个STR基因座的累积个体鉴别力(C_DP)为0.99999，累积非父排除能力(C_PE)为0.99578。6个STR基因座具有高度遗传多态性，可用于牛的遗传连锁分析、个体识别和亲权鉴定等研究领域。 2 应用微卫星分子标记对奶牛和肉牛进行亲权鉴定 运用PCR技术对BM2113、BM1862、BMc701、BM2934、TGLA122、BM720共6个STR基因座进行扩增，扩增产物分两组混合后经聚丙烯酰胺凝胶电泳和银染检测，扩增片段互不干扰。通过分析确定母女关系的2头奶牛血液样本和5个嫌疑父亲的精液样本的DNA，一头母本已知的奶牛在嫌疑父本中找出父亲，并计算父权概率，这6个STR基因座用于亲权鉴定获得99.993％的父权概率。证明用银染法检测STR基因座扩增产物可用于奶牛亲权鉴定。更多还原

【Abstract】 Genetic markers have been an absolutely necessarily research means since modern genetic has been established this century. STR locus complies Mendelian inheritance principles and distributes abroad and amplifies with PCR and polyacrylamide gel analyzed with sliver staining, so it is applied in molecular genetic science fields and developed startling. Individual identification and parentage tests are improved fast using microsatellite markers. The aims of the present study were:(1)to study the polymorphism distributions of six STR loci in dairy cattle and beef cattle;(2)to establish a nicety and shortcut method for and parentage tests in dairy cattle and beef cattle.1 The polymorphism distributions of STR loci in dairy cattle and beef cattleCatties for experiments were from two breeds, including 9 Holstein Friesian bull tissue samples and 49 Holstein Friesian cow blood samples and 103 beef cattle tissue samples. Extracted DNA from these samples and analyzed the polymorphism distributions of six STR loci in dairy cattle and beef cattle. The polymorphism distributions of six STR loci, BM2113, BM1862, BMc701, BM2934, TGLA122, BM720 were detected in cattle by Polymerase Chain Reaction multiplex gel electrophoresis followed by silver staining. Gene frequency (Pi) , power of discrimination (DP), heterozygosity (H), polymorphism information content (PIC) and probability of paternity exclusion (PE) were calculated. All loci meet Hardy-Weinberg equilibrium. DP, H and PIC of BM2113 locus, and PE of TGLA122 locus are the biggest among six STR loci in Holstein Friesian. Cumulate DP of six STR loci is 0.99997, Cumulate PE of six STR loci is 0.98827. DP, H, PIC and PE of BM1862 loci are the biggest among six STR loci in beef. Cumulate DP of six STR loci is 0.99999, Cumulate PE of six STR loci is 0.99578. The six STR loci have high genetic polymorphism and they could be used as linkage analysis, individual identification and paternity test in cattle.2 Application of STR Loci by Silver Staining And PCR Methods for Parentage Identification in Dairy Cattle and Beef cattleAmplifications of six STR loci such as BM2113, BM1862, BMc701, BM2934, TGLA 122, BM720 were performed by PCR (polymerase chain reaction) method. The products of amplification were separated in polyacrylamide gel and analyzed with sliver staining. The method used in the case could provide a power of paternity of approximately 99.993%. The results identifying a heifer’s paternity with known maternity showed that the sliver staining was a validation method for identification parentage in dairy cattle.更多还原