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

基本信息

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

项目摘要

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.

我们已经通过使用聚合酶链反应（PCR）（PCR）研究了免疫球蛋白G（IgG）和免疫球蛋白KAPPA轻链（IG KAPPA）基因的遗传多态性（IG KAPPA）。免疫球蛋白基因内的多态区域的扩增。从第一个PCR获得的扩增产物的小部分进一步受到第二个PCR扩增，其中使用了等位基因特异性引物组，其中用于在这种方法的基因分型中。 IgG3常数基因的等位基因G3M*T和NON-G3M*T等位基因，该等位基因确定G3M（T）同种型及其对立的异型非型非G3M（T），以及Ig Kappa常数基因，Km*1 km*1 km*1检查了基因分型的，检查了基因分型。对于Ig Kappa常数基因，353bp片段包含KM同型的多态性区域的353bp片段通过第一个PCR成功从样品DNA中扩增。使用第一个PCR的产品作为样品的第二个PCR可以清楚地识别三个等位基因之间的序列变化。在冈山县的72位个体中进行的一项少量人口研究的结果表明，基因频率为km*3 = 0.736和km*1.2 = 0.264.在检查IgG3常数基因的情况下，726bp片段包含包含多晶的多态片段G3M（T）同种型通过第一个PCR成功从样品DNA中扩增。使用第一个PCR的产物作为样品，第二个PCR通常可以区分两个等位基因之间的核苷酸的替换。