Establishment of a high throughput single-nucleotide-polymorphisms (SNPs) screening system based on the re-annealing and digestion reaction
基于复性和消化反应的高通量单核苷酸多态性(SNP)筛选体系的建立
基本信息
- 批准号:15590592
- 负责人:
- 金额:$ 2.37万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:2003
- 资助国家:日本
- 起止时间:2003 至 2006
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Se blood group is important in the forensic field because it is associated with the secretion of ABO blood group antigens in body fluids. In this study, first, Se1, Se2, sej and se alleles of the Se blood group which are common in Japanese population were genotyped using the temperature-gradient gel electrophoresis method. Heteroduplex bands occurred in the heterozygotic samples showed genotype-specific electrophoresis patterns. When one mismatched nucleotide existed, one or two heteroduplex bands were found in the gel. In the samples with two mismatched bases, four heteroduplex bands were observed. These findings suggested that numbers of the mismatched nucleotides could be estimated even when an unknown allele was analyzed by the method.This method was applied to the genotyping of ABO blood group. When exons 6 and 7 of the four typical alleles, A, B, O^<A-> and 0G were analyzed, the homoduplex bands showed allele-specific electrophoresis patterns due to difference in the GC contents. Different heteroduplex bands could be detected after mixture of different alleles, so that ABO blood group could be genotyped by this method.This method was also applied to identify the htpervariable region 1 of mitochondrial DNA (mtDNA HV1). Homoduplex bands of mtDNA HV1 showed different electrophoresis patterns, and mixture of different samples revealed specific heteroduplex bands, so that mtDNA HV1 variants could be identified by this method.Mismatched nucleotides in heteroduplex bands could not be cleaved by means of the reported techniques. So, the temperature-gradient gel electrophoresis was the most suitable, high throughput screening method to detect single nucleotide polymorphisms (SNPs) in alleles and variants.
Se血型在法医学领域具有重要意义,因为它与体液中ABO血型抗原的分泌有关。在这项研究中,首先,Se 1,Se 2,sej和se等位基因的Se血型,这是常见的日本人口的基因型,使用温度梯度凝胶电泳方法。杂合样本中出现的异源双链带显示出基因型特异的电泳图谱。当存在一个错配核苷酸时,在凝胶中发现一条或两条异源双链带。在有两个碱基错配的样品中,观察到四条异源双链带。这些结果表明,即使在分析未知等位基因时,该方法也可以估计错配核苷酸的数量。分析A、B、O^和O G四个典型等位基因的第6和第7外显子时<A->,由于GC含量的差异,同源双链带显示等位基因特异性电泳图谱。不同等位基因混合后可检测到不同的异源双链带,从而可用于ABO血型的基因分型,并将该方法应用于线粒体DNA高变区1(mtDNAHV1)的鉴定。mtDNA HV1的同源双链带显示不同的电泳图谱,不同样品的混合物显示特异的异源双链带,因此该方法可用于mtDNA HV1变异体的鉴定,而异源双链带中的错配核苷酸不能用文献报道的方法切割。因此,温度梯度凝胶电泳是检测等位基因和变异体单核苷酸多态性(SNPs)的最合适的高通量筛选方法。
项目成果
期刊论文数量(37)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
三谷友亮: "温度勾配電気泳動法によるFUT2遺伝子のSNP解析"DNA多型. 12巻(印刷中). (2004)
Yusuke Mitani:“通过温度梯度电泳进行 FUT2 基因的 SNP 分析”DNA 多态性第 12 卷(出版中)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
Analysis of SNP in FUT2gene by temperature gradient gel electrophoresis.
温度梯度凝胶电泳分析FUT2基因中的SNP。
- DOI:
- 发表时间:2004
- 期刊:
- 影响因子:0
- 作者:Mitani T;Akane;A;Kobayashi T;Tokiyasu T;Yoshimura S;Okii Y;Yoshida M;Watabiki T
- 通讯作者:Watabiki T
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AKANE Atsushi其他文献
AKANE Atsushi的其他文献
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{{ truncateString('AKANE Atsushi', 18)}}的其他基金
Investigation of novel methods detecting separated polymorphic nucleotide variations in blood group genes based on the inverse-PCR technique
基于反向PCR技术检测血型基因多态性核苷酸变异的新方法研究
- 批准号:
11670428 - 财政年份:1999
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Establishment of MNSs haplotyping methods by gene analytical techniques.
利用基因分析技术建立MNS单倍型分析方法。
- 批准号:
09670457 - 财政年份:1997
- 资助金额:
$ 2.37万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
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ABO血型不相容肾移植的免疫学风险效益分析和抗体治疗进展
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16K11063 - 财政年份:2016
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26293161 - 财政年份:2014
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Development of novel ABO blood group antigen targeting peptide that suppress rejection during ABO-incompatible kidney transplantation
开发新型 ABO 血型抗原靶向肽,可抑制 ABO 不相容肾移植过程中的排斥反应
- 批准号:
23791736 - 财政年份:2011
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$ 2.37万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
Elucidation of ABO blood group gene expression though chromatin remodeling
通过染色质重塑阐明 ABO 血型基因表达
- 批准号:
22390140 - 财政年份:2010
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Grant-in-Aid for Scientific Research (B)
Red Cell Transfusion and In-hospital Mortality: Impact of Age of Red Cells and ABO Blood Group
红细胞输注和院内死亡率:红细胞年龄和 ABO 血型的影响
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181862 - 财政年份:2009
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Operating Grants
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开发表达人ABO血型抗原的克隆猪耐受诱导方法
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21380168 - 财政年份:2009
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17209044 - 财政年份:2005
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Development of model animal to reveal the antibody production for ABO blood group antigens.
开发模型动物以揭示 ABO 血型抗原的抗体产生。
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Grant-in-Aid for Scientific Research (C)
Progress in the biological science of ABO blood group system -a pioneer work in legal medicine-
ABO血型系统生物科学进展-法医学的开创性工作-
- 批准号:
13307018 - 财政年份:2001
- 资助金额:
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Grant-in-Aid for Scientific Research (A)
Distribution of ABO blood group H type 1-4 sugar chain antigens in human tissue from secretor or non-secretor individuals
分泌者或非分泌者个体的人体组织中 ABO 血型 H 1-4 型糖链抗原的分布
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