法医学杂志

法医学杂志 ›› 2006, Vol. 0 ›› Issue (3): 193-195.

• 论文 • 上一篇    下一篇

PCR-RFLP法分析北方汉族人群HLA-B基因多态性

曾昭书;丁梅;朱运良;刘志芳;孙志刚;黄洪武;   

  1. 郑州大学基础医学院法医学教研室,中国医科大学法医学院血清室,郑州大学基础医学院法医学教研室,中国医科大学法医学院血清室,中国医科大学法医学院血清室,中国医科大学法医学院血清室 河南郑州450052中国医科大学法医学院血清室,辽宁沈阳110001,辽宁沈阳110001,河南郑州450052,辽宁沈阳110001,辽宁沈阳110001,辽宁沈阳110001
  • 发布日期:2006-06-25 出版日期:2006-06-28

A PCR-RFLP Analysis to HLA-B Gene among Chinese Northern Han Populations

ZENG ZHAO-SHU1,2, DING MEI2, ZHU YUN-LIANG1, LIU ZHI-FANG2, SUN ZHI-GANG2, HUANG HONG-WU2(1. DEPARTMENT OF FORENSIC MEDICINE, SCHOOL OF BASIC MEDICINE, ZHENGZHOU UNIVERSITY, ZHENGZHOU 450052,CHINA; 2. DEPARTMENT OF FORENSIC SEROLOGY, SCHOOL OF FORENSIC ME   

  • Online:2006-06-25 Published:2006-06-28

摘要： 目的建立以PCR-RFLP法分析HLA-B基因多态性的方法,并对105名北方汉族人群酶切片段类型、频率进行调查,为其法医学应用奠定基础。方法酚/氯仿抽提法抽提样本DNA;PCR扩增HLA-B基因的外显子2,3;NlaIII酶切PCR产物;聚丙烯酰胺凝胶电泳和银染技术检测酶切结果;DNA测序确认酶切位点。结果HLA-B特异性扩增片段长度为943bp,以NlaIII酶进行酶切时在北方汉族人群中获得了14条片段,20种谱带类型,观察到了6个酶切位点。结论仅应用一种内切酶对HLA-B基因进行PCR-RFLP分析即可获得多条片段,多态信息含量大大提高,该法可以应用于法医学亲子鉴定和个人识别等。

关键词: HLA-B基因, 限制性片段长度多态性, 个人识别力

Abstract: Objective To set up the method for analyzing HLA-B gene polymorphism with PCR-RFLP, and to gain population data among northern Chinese Hans of HLA-B′s restricted fragments after NlaIII digestion, and to achieve application in forensic medicine practice. Methods Sample DNA was extracted by the phenol/chloroform extraction method, 943 bp-long fragments containing HLA-B exon 2 and 3 were got by PCR. The endonuclease NlaIII was applied to cut the PCR products into polymorphic fragments shorter than 943bp, then PAGE and silver staining were used to detect the digestion results, finally the digestion sites were assured by DNA sequencing. Results Along 943bp-long PCR products, 14 length-different fragments, 20 kinds of fragment combinations were got and 6 cutting site were observed after NlaIII digestion. Conclusion HLA-B gene was highly polymorphic among Chinese northern Hans. Even with only one endonuclease, 14 restricted fragments were got and the PIC was great. Such a HLA-B PCR-RFLP analysis will have values in forensic medicine applications.

Key words: HLA-B gene, restriction fragment length polymorphism, discrimination power