Gm genotyping from forensic biological materials by DNA hybridization

通过 DNA 杂交对法医生物材料进行转基因基因分型

• 批准号: 03670297
• 负责人: ISHIZU Hideo
• 金额: $ 1.28万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670297/
• 关键词: Immunoglobulin; Genetic polymorphism; Gm type; Km type; DNA; Polymerase chain reaction; 免疫グロブリンG; ポリメラ-ゼ・チェ-ン・リアクション

We have investigated the genetic polymorphisms of immunoglobulin G (IgG) and immunoglobulin kappa light chain (Ig kappa) genes by using the polymerase chain reaction (PCR).In the present methods, samples of extracted human genome DNA were subjected to the first PCR for the amplification of the polymorphic regions within the immunoglobulin genes.Small portions of the amplification products obtained from the first PCR were further subjected to the second PCR amplifications in which the allele specific primer sets were used for genotyping of the immunoglobulin genes.By this methods, alleles on the IgG3 constant gene, G3m*t andnon-G3m*t alleles, which determine the G3m(t) allotype and its antithetical isoallotype non-G3m(t), respectively, and alleles on the Ig kappa constant gene, Km*1, Km*1.2 and Km*3, were examined for genotyping.For the examination of the Ig kappa constant gene, 353bp fragments which contain the polymorphic region of the Km allotypes were successfully amplified from sample DNAs by the first PCR. The second PCRs using the products of first PCR as samples could clearly identify the sequence changes between the three alleles. Results of a tiny population study among 72 individuals living in Okayama Prefecture showed the gene frequencies of Km*3= 0.736 and Km*1.2= 0.264.In the case of the examination of the IgG3 constant gene, 726bp fragments which contain the polymorphic region of the G3m(t) allotype were successfully amplified from sample DNAs by the first PCR. Using the products of first PCR as samples, the second PCRs could generally distinguish the nucleotide substision between the two alleles.

我们调查了免疫球蛋白G（IgG）和免疫球蛋白κ轻链的遗传多态性，（IG kappa）基因。在本方法中，对提取的人类基因组DNA样品进行第一次PCR，以扩增免疫球蛋白基因内的多态性区域。通过该方法，IgG 3恒定基因上的等位基因，G3m*t和非G3m *t等位基因，其分别决定G3m（t）同种异型和其对立同种异型非G3m（t），以及IG κ恒定基因上的等位基因，Km*1，Km*1.2和Km*3，第一次PCR成功扩增出353 bp的Km等位基因多态性片段，用于IG κ恒定区基因的检测。以第一次PCR产物为样本的第二次PCR可以清楚地识别出三个等位基因之间的序列变化。对冈山县72个个体的小群体研究结果表明，基因频率Km*3= 0.736，Km*1.2= 0.264。在IgG3恒定基因检测中，首次PCR成功地从样本DNA中扩增出含有G3m（t）同种异型多态性区域的726bp片段。以第一次PCR产物为样本，第二次PCR基本可以区分两个等位基因之间的核苷酸替换。