Investigation of mechanism for the genesis of polymorphisms of the coagulation factor XIII and PGM1 genes

凝血因子XIII和PGM1基因多态性发生机制的研究

• 批准号: 05670403
• 负责人: SUZUKI Koichi
• 金额: $ 1.02万
• 依托单位: Osaka Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670403/
• 关键词: coagulation factor XIII; genetic polymorphism; base substitution; haplotype; rare variant; factor XIII deficiency; recombination hot-spot; 凝固第13因子; 凝固因子

We demonstrated molecular basis of the genetic polymorphism of the coagulation factor XIII "a" subunit and novel sequence polymorphisms in five exons. Four F13A alleles were found to be defined by nucleotide substitutions at the three sites of exon 12 and exon 14. We developed PCR-RFLPs by which these substitutions can be detected at the genomic level. In addition to the three sites, we characterized novel polymorphic sites of exons 2,5,8,12 and 14 in Caucasian populations whereas these sites were non-polymorphic in Japanese except for the site of exon 2. These polymorphic sites were found to be transmitted as haplotypes. A total of 18 haplotypes out of theoretically 72 possible combinations were found to occur in the Caucasian populations, implying that frequent recombination events rather than reccurent point mutations lead to production of the haplotypes. Furthermore, we revealed sequence differences for 14 distinct variant alleles and one deficient allele by characterizing their complete sequences. Amino acid substitutions resulting from the nucleotide changes of the 14 variant alleles were consistent with mobility shift of their products in IEF gels. The deficient allele was found to result from abnormal splicing caused by a single nucleotide mutation at the splice-acceptor site in intron 5 of the F13A gene.The data obtained in this study suggest that there are recombination hot-spots in the F13A gene region. We are now investigating the sequences of introns and 5'-and 3'-flanking regions.

我们展示了凝血因子XIII“a”亚基遗传多态的分子基础和5个外显子上新的序列多态。发现4个F13A等位基因是由外显子12和外显子14的三个位点的核苷酸替换所定义的。我们建立了在基因组水平上检测这些替换的PCR-RFLP。除这三个外显子外，我们还在高加索人群中发现了新的外显子2、5、8、12和14的多态位点，而在日本人中，除了外显子2外，这些新的多态位点都是非多态的。这些多态位点被发现是以单倍型传递的。在理论上72个可能的组合中，共有18个单倍型出现在高加索人群中，这意味着导致单倍型产生的是频繁的重组事件，而不是复发性的点突变。此外，我们还通过对14个不同的变异等位基因和1个缺失等位基因的全序列特征分析，揭示了它们的序列差异。14个变异等位基因的核苷酸变化导致的氨基酸替换与其产物在IEF凝胶中的迁移率移动是一致的。该缺失等位基因是由于F13A基因第5内含子剪接受体处的单核苷酸突变引起的异常剪接所致，提示F13A基因区域存在重组热点。我们现在正在研究内含子以及5‘和3’侧翼区的序列。

项目成果

K.Suzuki et al.: "Molecular basis for polymorphism of the "a"subunit of coagulation factor XIII." Advances in Forensic Haemogenetics. 5. 424-426 (1994)

K.Suzuki 等人：“凝血因子 XIII 的“a”亚基多态性的分子基础。”

Suzuki,et al: "A study on ADH2 and ALDH2 genotyping by PCR-RFLP and SSCP analyses with description of allele and genotype frequencies in Japanese,Finn,and Lapp populations." Alcohol & Alcoholism. 29(sup1.),(in print). (1995)

Suzuki 等人：“通过 PCR-RFLP 和 SSCP 分析进行 ADH2 和 ALDH2 基因分型的研究，并描述了日本人、芬兰人和拉普人人群中的等位基因和基因型频率。”

鈴木広一 他: "凝固第XIII因子aサブユニット遺伝子に見いだされた新たな多型について" DNA多型. 3(印刷中). (1995)

Koichi Suzuki 等人：“凝血因子 XIII 亚基基因中发现的新多态性”DNA 多态性 3（出版中）。

Suzuki,et al: "DNA typing of coagulation factor XIII “a" subunit by PCR-RFLP and SSCP." Advances in Forensic Haemogenetics. 5. 173-175 (1994)

Suzuki 等人：“通过 PCR-RFLP 和 SSCP 对凝血因子 XIII“a”亚基进行 DNA 分型。法医血液遗传学进展”5. 173-175 (1994)

Koichi Suzuki, Misa Iwata, Shigenori Ito, Kiyoshi Matsui, Atsuko Uchida, Yasuhiko Mizoi.: "Molecular basis for subtypic differences of the "a" subunit of coagulation factor XIII with description of the genesis of the subtypes." Human Genetics. 94. 129-135

Koichi Suzuki、Misa Iwata、Shigenori Ito、Kiyoshi Matsui、Atsuko Uchida、Yasuhiko Mizoi.：“凝血因子 XIII 的“a”亚基亚型差异的分子基础以及亚型起源的描述。”