【作者】 夏水秀；

【导师】 冯一中；

【作者基本信息】 苏州大学 ， 病理学， 2007， 硕士

【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。更多还原

【Abstract】 Objective The aim of this study is to obtain the genetic polymorphisms data of D2S1399 and D5S2500 STR loci in Eastern Chinese Han population, and evaluate the forensic application of the two loci. The standard allelic ladders for D2S1399 and D5S2500 STR loci were constructed by molecular clone.Methods Blood samples or buccal swabs of unrelated Han individuals living in eastern China were analyzed using PCR-nature polyacrylamide gel electrophoresis- sliver staining method. Different PCR amplified D2S1399 and D5S2500 loci allelic fragments were isolated from the polyacrylamide gel. The STR allelic fragments were eluted into distilled water and reamplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors. The size and structure of the inserts were confirmed by the analysis of their DNA sequences. The PUC plasmid vectors were transfected into E. coli DH5αTM cells, which were cultivated on selective media. The recombinant plasmids DNA with the inserts were then used as template for reamplification to generate D2S1399 and D5S2500 loci standard ladders.Results 11 alleles of D2S1399 locus and 9 alleles of D5S2500 locus were observed in Eastern Chinese Han population respectively. The observed heterozygosity (Ho) values, the discrimination power (DP) values and the power of exclusion values of D2S1399 and D5S2500 were 0.745 and 0.807, 0.958 and 0.917, 0.554 and 0.643 , respectively. A large quantity of standard allelic ladder for D2S1399 and D5S2500 were obtained.Conclusion D2S1399 and D5S2500 loci were highly informative and suitable for forensic application. It’s feasible to construct the standard allelic ladder of D2S1399 and D5S2500 loci for STR typing by allele molecular cloning technic.更多还原