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Analysis of cystic fibrosis transmembrane conductance regulator gene in patients with diffuse panbronchiolitis.

弥漫性全细支气管炎患者囊性纤维化跨膜电导调节基因分析

基本信息

批准号：
03454235
负责人：
NUKIWA Toshihiro
金额：
$ 2.75万
依托单位：
Juntendo University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
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/en/grant/KAKENHI-PROJECT-03454235/
关键词：
Respiratory epithelial cell CFTR gene Chloride ion channel Cystic fibrosis Diffuse panbronchiolitis SSCP

项目摘要

Diffuse panbronchiolitis (DPB), relatively popular among non-smoker adults in Japan, is characterized by chronic inflammation located in respiratory bronchioles, causing obstructive respiratory failure with persistent pseudomonas infection, and frequent complication with chronic sinusitis. Recent elucidation of the genetic defect in a putative chloride ion channel gene, cystic fibrosis transmembrane conductance regulator (CFTR), among patients with cystic fibrosis (CF) who shares resembling clinical features with DPB led us to examine possible genetic defects in CFTR gene among patients with DPB in Japan. The Cystic Fibrosis Genetic Analysis Consortium accumulated and reported almost 300 sequence alterations in the CFTR gene, frequently associated with exons consisting peptides of putative functional domains of CFTR protein. In this context, we examined, in the 1st year, exons 4,7,10,11,19,20 among 27 exons in CFTR gene where genetic defects are highly accumulated, and then extended to … More all exons in the 2nd year. DNA fragments were amplified by polymerase chain reaction using genomic DNAs from 18 patients with DPB (age 18 - 74; 11 males, 7 females). The fragments were then subjected to electrophoresis for single strand conformational polymorphism (SSCP). In exons 12 and 18, two different SSCPs' were found in 3 cases, and in 3 and 24, single SSCP was found in 2 cases, respectively. In exons 9,11,19,21,22, and 24, single SSCP was detected in one case. In exons 6b,9,10, and 24, there found SSCPs' with patterns of homozygous and heterozygous bands, suggesting the existence of normal variant polymorphism. In exons 2,4,5,7,8,13,14a,14b,15,17b,20, and 23, no SSCP was detected in all 18 cases. Exons 1,13,16,and 17a should be analyzed by direct sequencing. These results indicate that the genetic defects in patients with DPB in Japan may not be related to the CFTR gene, because no common SSCP bands was found among 18 patients, although direct sequencing of all CFTR exons in one or two patients is necessary to neglect false negatives due to SSCP procedure. Less

弥漫性泛细支气管炎（diffusepanbronchiolitis，DPB）在日本不吸烟的成年人中较为常见，其特征是位于呼吸性细支气管的慢性炎症，导致阻塞性呼吸衰竭伴持续性假性感染，并经常并发慢性鼻窦炎。最近阐明的一个假定的氯离子通道基因，囊性纤维化跨膜传导调节因子（CFTR）的遗传缺陷，囊性纤维化（CF）的患者与DPB相似的临床特征，导致我们检查可能的遗传缺陷，在日本的DPB患者的CFTR基因。囊性纤维化遗传分析联盟积累并报道了CFTR基因中近300个序列改变，这些改变通常与构成CFTR蛋白推定功能结构域的肽的外显子相关。在此背景下，我们在第一年检查了CFTR基因中遗传缺陷高度累积的27个外显子中的第4、7、10、11、19、20个外显子，然后扩展到 ...更多信息第二年的所有外显子。使用来自18名DPB患者（年龄18 - 74岁; 11名男性，7名女性）的基因组DNA，通过聚合酶链反应扩增DNA片段。然后对片段进行单链构象多态性（SSCP）电泳。在第12和18外显子中，3例出现两种不同的SSCP，在第3和24外显子中，分别有2例出现单一的SSCP。在第9、11、19、21、22和24外显子中，有1例检测到单一SSCP。在外显子6 b、9、10和24中发现了SSCP的纯合和杂合带型，提示存在正常的变异多态性。在外显子2、4、5、7、8、13、14 a、14 b、15、17 b、20和23中均未检测到SSCP。外显子1、13、16和17 a应通过直接测序进行分析。这些结果表明，日本DPB患者的遗传缺陷可能与CFTR基因无关，因为在18名患者中没有发现共同的SSCP条带，尽管有必要对1名或2名患者的所有CFTR外显子进行直接测序，以忽略SSCP程序导致的假阴性。少