Molecular cloning of a mouse DNA repair gene that complements the defect of group-A xeroderma pigmentosum.
For isolation of the gene responsible for xeroderma pigmentosum (XP) complementation group A, plasmid pSV2gpt and genomic DNA from a mouse embryo were cotransfected into XP2OSSV cells, a group-A XP cell line. Two primary UV-resistant XP transfectants were isolated from about 1.6 X 10(5) pSV2gpt-transformed XP colonies. pSV2gpt and genomic DNA from the primary transfectants were again cotransfected into XP2OSSV cells and a secondary UV-resistant XP transfectant was obtained by screening about 4.8 X 10(5) pSV2gpt-transformed XP colonies. The secondary transfectant retained fewer mouse repetitive sequences. A mouse gene that complements the defect of XP2OSSV cells was cloned into an EMBL3 vector from the genome of a secondary transfectant. Transfections of the cloned DNA also conferred UV resistance on another group-A XP cell line but not on XP cell lines of group C, D, F, or G. Northern blot analysis of poly(A)+ RNA with a subfragment of cloned mouse DNA repair gene as the probe revealed that an approximately 1.0 kilobase mRNA was transcribed in the donor mouse embryo and secondary transfectant, and approximately 1.0- and approximately 1.3-kilobase mRNAs were transcribed in normal human cells, but none of these mRNAs was detected in three strains of group-A XP cells. These results suggest that the cloned DNA repair gene is specific for group-A XP and may be the mouse homologue of the group-A XP human gene.
Tanaka
K
K
Institute for Molecular and Cellular Biology, Osaka University, Japan.
Satokata
I
I
Ogita
Z
Z
Uchida
T
T
Okada
Y
Y
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000528:radiation effects; D003001:Cloning, Molecular; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects; D004622:Embryo, Mammalian; D005796:Genes; D005816:Genetic Complementation Test; D006801:Humans; D051379:Mice; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection; D014466:Ultraviolet Rays; D014983:Xeroderma Pigmentosum / Q000235:genetics*