Molecular analyses of UV^s syndrome and development of simple diagnostic methods for inherited photosensitive diseases

我利用这笔资金调查了以下实验。1)遗传性光敏疾病简单诊断方法的发展为了获得色素性干皮病A ~ G组（E组除外）和Cockayne综合征A、B组等光敏性疾病候选者，我采用集落杂交或PCR的方法进行了cdna的分离。将cdna亚克隆到真核表达载体（pcDNA1、pcDNA3、pTarget、pcDM8）中。3 .建立了利用3种DNA修复标记(UDS (unplanned DNA synthesis), RRS （recovery of RNAS synthesis), RDS (recovery of replication DNA synthesis)）对光敏性疾病进行分类的方法。根据分类，我们采用显微注射或转染方法建立了明确的诊断方法。2) uv ^ s综合征的分子闭合。利用SV40 T抗原2建立了两个独立的不灭UV^SS克隆（Kps3SVY和Kps3SVI3）。为了证实两个不朽克隆来源于亲代原代细胞，我们进行了VNTR（可变数目串联重复）分析。3 .利用HeLa S3 mRNA（大小分形（+））构建EB病毒cDNA文库。构建了表达EBNA抗原的不灭UV^SS克隆。我现在正在筛选补充UV^SS缺陷的候选基因。

I investigated the following experiments using this grant.1) DEVELOPMENT OF SIMPLE DIAGNOSTIC METHODS FOR INHERITED PHOTO-SENSITIVE DISEASES1. To obtain the candidates of photosensitive diseases, such as xeroderma pigmentosum group A through G (except for E) and Cockayne syndrome group A and B, I isolated the cDNAs using colony-hybridization or PCR method.2. cDNAs were subcloned into eukaryotic expression vectors (pcDNA1, pcDNA3, pTarget, pcDM8).3. I established the method of classification of photosensitive diseases using three DNA repair markers (UDS (unschduled DNA synthesis), RRS (recovery of RNAS synthesis), and RDS (recovery of replicative DNA synthesis)).4. Following the classification, we establised the definitive diagnostic method using micro-injection or transfection methods.2) MOLECULAR CLOING OF UV^S SYNDROME1. I established two independent immrotal UV^SS clones (Kps3SVY and Kps3SVI3) using SV40 T antigen.2. To confirm that two immrotal clones were derived from parental primary cells, we performed VNTR (variable number of tandem repeat) analyses.3. I constructed EB virus based cDNA library using HeLa S3 mRNA (size fractination (+)).4. I constructed immortal UV^SS clones expressed EBNA antigen.5. I am now screening the candidate gene complemented the UV^SS defect.