法医学杂志

法医学杂志 ›› 2001, Vol. 0 ›› Issue (3): 148-151.

• 论文 • 上一篇    下一篇

D1S549基因座分型标准物的克隆制备方法及其群体遗传学研究

张林,辛军平,陈国弟,田新,廖淼,李荣华   

  1. 四川大学华西法医学院!四川成都 610041,四川大学华西法医学院!四川成都 610041,四川大学华西法医学院!四川成都 610041,四川大学华西法医学院!四川成都 610041,四川大学华西法医学院!四川成都 610041,四川大学华西法医学院!四川成都 610041
  • 发布日期:2001-06-25 出版日期:2001-06-28

Study on the construction of standard D1S549 allelic ladder via molecular clonning and its genetic polymorphism in Chinese three populations

ZHANG LIN,XIN JUN PING,CHEN GUO DI,ET AL.(COLLEGE OF FORENSIC MEDICINE,SICHUAN UNIVERSITY,CHENGDU 610041)   

  • Online:2001-06-25 Published:2001-06-28

摘要： 目的 采用分子克隆技术建立制备大量优质标准 STR基因座等位基因分型标准物的方法，解决长期困扰 STR分型上存在的准确性和标准化问题。方法 先用 PCR扩增出 D1S549基因座的 8个等位基因片段，将其插入 pUC重组质粒中，经 DNA测序分析证实插入片段的结构及大小，用国际标准将插入的等位基因片段进行命名，最后经转染、扩大培养，扩增及再鉴定后制备出标准的 D1S549等位基因分型标准物。结果应用此分型标准物，调查了成都地区汉族 ,甘肃回族及新疆维族群体 D1S549基因座的基因型分布频率。结论 表明该法制备的 STR基因座等位基因分型标准物适用于法医学鉴定实践， D1S549基因座是一个适合我国群体分析和法医学遗传分析的遗传标记。

关键词: 等位基因分型标准物, 短串联重复序列, D1S549基因座, 遗传多态性

Abstract: Objective To resolve the problem of the accuracy and standardization of STR－ PCR typing in forensic practice, we have designed a new method to produce standard D1S549 allelic ladder. Methods Eight different PCR amplified D1S549 allelic fragments were isolated from the gel, eluted into the distilled water and re－ amplified by PCR. The purified allelic fragments were then blunt－ end subcloned individually into the pUC plasmid vectors and transfected into competent E.coli DH5α TM cells.Results The sequencing results confirmed that the size and the construe of the inserts were correct. The recombinant plasmids DNA with 8 inserts were then used as templates for re－ amplification to generate D1S549 standard ladder, with which the genetic polymorphisms of D1S549 locus in Chinese Han population in chengdu, Hui population in Gansu and Wei population in Xinjiang were studied. Conclusion The results showed that the standard ladder made via this method is excellent, and D1S549 locus is robust for genetic research and forensic application.

Key words: allelic ladder, short tandem repeat, D1S549 locus, genetic polymorphism