Molecular pathogenesis of the patients with congenital disorder of thrombosis and hemostasis

I. I. Analysis of Bernard-Soulier syndrome ： PheイD155エD1 （TTT） to Ser （TCT） replacement in the leucine rich motif （LRM） of the GPIX polypeptide does cause BSS phenotype。我们已通过对突变GPIX的体外转染研究和GPIb/IX复合体的其他等离子体进行体外转染研究的含义进行认证。突变GPIX不能增加GPIb的表面表达--GPIX本身的非表面表达。Analysis of coagulation factor X deficiency ： We have identified the molecular defect underlying congenital factor X （FX） deficiency。一部小说3-基对删除被发现在FX基因的Intron D中。在53个不相关的日文（106个段落）中没有检测到这一突变，通过一个特定的限制分析，表明了外汇缺陷的可能原因。The Deletion resides within a polypyrimidine tract of the acceptor splicing site where U2snRNP binds to form spliceosomes.这一缺陷可能改变胚胎的形成，然后在胚胎和退化中的结果不正确 ... More ased FX production. III。对协同系数XI缺陷的分析：我们已经确定了一个新颖的单基点突变，一个GT->因子XI （FXI）基因的转变在其内含子J. A反向转录聚合酶链反应，该反应显示了在FXI exon 10的3'-end的20基转录，在正确的前10/11交叉点10/11交叉点56个下游的框架移位和生成中的结果。这种突变的一种现象性后果，然后通过哺乳动物细胞表达系统进行了调查。再组合的等离子体生长野生型FXI基因直接将FXI抗原的生产转化为中间体，但在FXI exon 10中丢失的20个等离子体，在mRNA表达未改变的情况下生产抗原。这里描述的突变是一种新颖的、来自外围单核细胞的电子RNA，首次被提供用于分析结果FXI mRNA序列。Less（低）

I. Analysis of Bernard-Soulier syndrome : PheィイD155ィエD1 (TTT) to Ser (TCT) replacement in the leucine rich motif (LRM) of the GPIX polypeptide does cause BSS phenotype. We have certified this by means of in vitro transfection studies with plasmid for mutant GPIX and other plasmids for GPIb/IX complex. Mutant GPIX could not increase the surface expression of GPIb-α nor surface expression of GPIX itself.II. Analysis of coagulation factor X deficiency : We have identified the molecular defect underlying congenital factor X (FX) deficiency. A novel 3-base-pair deletion was observed within intron D of the FX gene. This mutation was not detected in 53 unrelated Japanese (106 alleles) by an allele-specific restriction analysis, indicating a likely cause of the FX deficiency. The deletion resides within a polypyrimidine tract of the acceptor splicing site where U2 snRNP binds to form spliceosomes. This defect could alter the formation of splicesomes, then result in incorrect splicing and decre … More ased FX production.III. Analysis of coagulation factor XI deficiency : We have identified a novel single-base point mutation, a GT->AT transition of the factor XI (FXI) gene at the donor splicing site of its intron J.A reverse transcription-polymerase chain reaction with ectopic RNA revealed that propositus cDNA showed 20-base truncation of the 3'-end of the FXI exon 10, resulting in the frame-shift and generation of premature stop codon at 56-base downstream of the correct exon 10/11 junction. A phenotypic consequence of this mutation was then investigated through mammalian cell expression system. Recombinant plasmids harboring wild type FXI gene did direct the production of the FXI antigen into the medium, but the plasmids lacking 20 bases of the FXI exon 10 failed to produce the antigen while the mRNA expression thereof was unchanged. The mutation described here is novel, and ectopic RNA from peripheral mononuclear cells was first proved to be useful for analyzing the resultant FXI mRNA sequence. Less