Zhang Z P, Blombäck M, Egberg N, Falk G, Anvret M
Department of Clinical Genetics, Karolinska Hospital, Stockholm, Sweden.
Genomics. 1994 May 1;21(1):188-93. doi: 10.1006/geno.1994.1241.
Twenty-four patients with von Willebrand disease type III were screened for mutations in the von Willebrand factor (VWF) gene using the PCR technique, followed by direct sequencing. More than 250 kb of genomic DNA were sequenced, including the promoter and coding regions (52 exons) of the VWF gene from 24 patients. In addition to the previously reported mutations of a single cytosine deletion in exon 18 and the nonsense mutations in exons 28, 32, and 45, nine new mutations were detected: two nonsense mutations in exons 15 and 16, one allele with a thymidine insertion in exon 14, one allele with a cytosine insertion in exon 28, one 20-bp deletion in exon 15, one mutation in the donor splice site of exon 43, and three missense mutations in exons 28, 49, and 51. Forty-two mutant chromosomes were identified (42/48); 11 probands are homozygous for the mutations, and 8 are compound heterozygous. In addition, a new subfamily of the Alu sequence in the promoter region and 10 new polymorphisms were identified.
采用聚合酶链反应(PCR)技术对24例Ⅲ型血管性血友病患者的血管性血友病因子(VWF)基因进行突变筛查,随后进行直接测序。对超过250kb的基因组DNA进行了测序,包括24例患者VWF基因的启动子和编码区(52个外显子)。除了先前报道的外显子18中单个胞嘧啶缺失突变以及外显子28、32和45中的无义突变外,还检测到9个新突变:外显子15和16中的两个无义突变、外显子14中一个等位基因的胸腺嘧啶插入、外显子28中一个等位基因的胞嘧啶插入、外显子15中的一个20bp缺失、外显子43供体剪接位点的一个突变以及外显子28、49和51中的三个错义突变。共鉴定出42条突变染色体(42/48);11名先证者为突变纯合子,8名为复合杂合子。此外,在启动子区域鉴定出一个新的Alu序列亚家族和10个新的多态性。