【摘要】 目的短串联重复序列(shorttandemrepeats,STR)分型的准确性和标准性是目前急需解决的关键问题之一,本研究采用分子克隆技术制备优质标准的4个(D1S1676、D2S2735、D11S1977和D22S444)STR基因座等位基因分型标准物,并用于中国成都汉族人群的群体研究,探讨该技术的实用性。方法PCR扩增出4个基因座的各等位基因片段后,将其插入pGEMR-T重组质粒中,DNA测序证实插入片段的大小和结构,等位基因片段按国际标准进行命名后,经转染、扩大培养、扩增及再鉴定后制备出该4个基因座的等位基因分型标准物,进行群体研究。结果这4个基因座的等位基因分型标准物得以大量成功制备,并成功地应用于成都地区汉族群体研究。结论分子克隆技术是制备STR等位基因分型标准物的可靠方法;群体资料显示,D1S1676和D2S2735基因座是一个适合中国成都汉族群体分析和法医学应用的遗传标记,而D11S1977和D22S444基因座在中国成都汉族群体中的应用价值有限。更多还原

【Abstract】 Objective To solve the problems in the accuracy and standardization of short tandem repeats-polymerase chain reaction (STR-PCR) typing, the authors adopted the molecular clone technology in producing the standard allelic ladders of D1S1676, D2S2735, D11S1977 and D22S444 loci and applied them in a population study on the Hans in Chengdu, China. Methods PCR was used to produce several different allelic fragments of these loci. PCR products were eluted from the gel and re-amplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pGEMR-T plasmid vectors and the recombinant were transfected into competent E.coli DH5α TM cells. The results of sequencing confirmed that the size and the construction of the inserts were correct. The recombinant plasmids DNA with the inserts were then used as template for re-amplification to generate the four loci standard ladders. Results The authors succeeded in producing large quantity of standard allelic ladder of these four loci, with which the genetic polymorphisms of these loci in Chengdu Han population of China were studied. Conclusion This method is of high value for forensic DNA typing to construct standard ladders. D1S1676,D2S2735 loci are robust for forensic analysis in Chinese Han population, whereas the value of D11S1977 and D22S444 loci is limited.更多还原