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

Research Project

Project/Area Number 09670887
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Dermatology
Research InstitutionKumamoto University

Principal Investigator

ITOH Toshiki  Kumamoto University School of Medicine, Research associate, 医学部, 助手 (00284753)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1997: ¥2,000,000 (Direct Cost: ¥2,000,000)
KeywordsUV^S syndrome / Xeroderma pigmentosum / Cockayne syndrome / UV^s症候群
Research Abstract

I investigated the following experiments using this grant.
1) DEVELOPMENT OF SIMPLE DIAGNOSTIC METHODS FOR INHERITED PHOTO-
SENSITIVE DISEASES
1. 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 SYNDROME
1. 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.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report

URL: 

Published: 1997-04-01   Modified: 2016-04-21  

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