1998 Fiscal Year Final Research Report Summary
Molecular analyses of UV^s syndrome and development of simple diagnostic methods for inherited photosensitive diseases
| Project/Area Number |
09670887
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | Kumamoto University |
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Principal Investigator |
ITOH Toshiki Kumamoto University School of Medicine, Research associate, 医学部, 助手 (00284753)
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| Project Period (FY) |
1997 – 1998
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| Keywords | UV^S syndrome / Xeroderma pigmentosum / Cockayne syndrome |
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| 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.
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