华东地区汉族人群D2S1399和D5S2500基因座的遗传多态性研究及其等位基因分型标准物的构建
发布时间：2018-10-26 19:59
【摘要】： 目的研究D2S1399和D5S2500基因座在中国华东地区汉族群体的遗传多态性,评估其在法科学中的应用价值;利用重组质粒制备这两个STR基因座的分型标准物。 方法应用PCR、聚丙烯酰胺垂直电泳及银染技术检测D2S1399和D5S2500基因座的遗传多态性。从聚丙烯酰胺凝胶中分离出经PCR扩增后的等位基因片段,二次扩增并纯化后的等位基因片段被分别插入到PUC质粒载体中。利用DNA测序证实插入片段大小及结构的正确性后,用国际标准将插入的等位基因片段进行命名。再转染到适当的E. coli DH5αTM细胞内选择培养,以纯化质粒为模板,扩增出各基因座的等位基因片段后混合获得等位基因分型标准物。 结果D2S1399和D5S2500基因座在中国华东汉族群体分别检出11个和9个等位基因,其观察杂合度分别为0.745和0.807,多态信息含量(PIC)分别为0.850和0.750,个体识别率分别为0.958和0.917,非父排除率(PE)分别为0.554和0.643。成功地建立了能大量生产的D2S1399及D5S2500基因座的标准等位基因片段的重组质粒库,并制备出大量优质分型标准物。 结论D2S1399和D5S2500基因座是高度多态性的STR基因座,在法医学个人识别和亲子鉴定中有重要应用价值。通过分子克隆制备D2S1399及D5S2500基因座的等位基因分型标准物是一种实现D2S1399及D5S2500基因座分型标准化的可行方法。
[Abstract]:Objective to study the genetic polymorphisms of D2S1399 and D5S2500 loci in the Han population of East China, to evaluate their application value in French science, and to prepare the standard compounds of the two STR loci by using recombinant plasmids. Methods PCR, polyacrylamide vertical electrophoresis and silver staining were used to detect the genetic polymorphism of D2S1399 and D5S2500 loci. The alleles amplified by PCR were isolated from the polyacrylamide gel and inserted into the PUC plasmid vector after secondary amplification and purification. After DNA sequencing was used to confirm the correctness of the size and structure of the inserted fragment, the inserted allele fragment was named by international standard. The alleles of each locus were amplified by using purified plasmids as template, and then the alleles were mixed to obtain the standard alleles. Results 11 and 9 alleles of D2S1399 and D5S2500 loci were detected in the Han population of East China respectively. The observed heterozygosity was 0.745 and 0.807, and the polymorphic information content (PIC) was 0.850 and 0.750, respectively. The individual recognition rates were 0.958 and 0.917, and the non-paternal exclusion rates (PE) were 0.554 and 0.643, respectively. A recombinant plasmid library of standard allelic fragments of D2S1399 and D5S2500 loci was successfully constructed and a large number of high quality genotyping standard materials were prepared. Conclusion D2S1399 and D5S2500 loci are highly polymorphic STR loci, which have important application value in forensic personal identification and paternity identification. The preparation of allelic alleles of D2S1399 and D5S2500 loci by molecular cloning is a feasible method for the standardization of D2S1399 and D5S2500 loci.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R394
本文编号：2296795
[Abstract]:Objective to study the genetic polymorphisms of D2S1399 and D5S2500 loci in the Han population of East China, to evaluate their application value in French science, and to prepare the standard compounds of the two STR loci by using recombinant plasmids. Methods PCR, polyacrylamide vertical electrophoresis and silver staining were used to detect the genetic polymorphism of D2S1399 and D5S2500 loci. The alleles amplified by PCR were isolated from the polyacrylamide gel and inserted into the PUC plasmid vector after secondary amplification and purification. After DNA sequencing was used to confirm the correctness of the size and structure of the inserted fragment, the inserted allele fragment was named by international standard. The alleles of each locus were amplified by using purified plasmids as template, and then the alleles were mixed to obtain the standard alleles. Results 11 and 9 alleles of D2S1399 and D5S2500 loci were detected in the Han population of East China respectively. The observed heterozygosity was 0.745 and 0.807, and the polymorphic information content (PIC) was 0.850 and 0.750, respectively. The individual recognition rates were 0.958 and 0.917, and the non-paternal exclusion rates (PE) were 0.554 and 0.643, respectively. A recombinant plasmid library of standard allelic fragments of D2S1399 and D5S2500 loci was successfully constructed and a large number of high quality genotyping standard materials were prepared. Conclusion D2S1399 and D5S2500 loci are highly polymorphic STR loci, which have important application value in forensic personal identification and paternity identification. The preparation of allelic alleles of D2S1399 and D5S2500 loci by molecular cloning is a feasible method for the standardization of D2S1399 and D5S2500 loci.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R394
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1 夏水秀;华东地区汉族人群D2S1399和D5S2500基因座的遗传多态性研究及其等位基因分型标准物的构建[D];苏州大学;2007年
，本文编号：2296795
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