Isolation and characterization of the human uracil DNA glycosylase gene
- Temple Univ. School of Medicine, Philadelphia, PA (USA)
A series of anti-human placental uracil DNA glycosylase monoclonal antibodies was used to screen a human placental cDNA library in phage {lambda}gt11. Twenty-seven immunopositive plaques were detected and purified. One clone containing a 1.2-kilobase (kb) human cDNA insert was chosen for further study by insertion into pUC8. The resultant recombinant plasmid selected by hybridization a human placental mRNA that encoded a 37-kDa polypeptide. This protein was immunoprecipitated specifically by an anti-human placenta uracil DNA glycosylase monoclonal antibody. RNA blot-hybridization (Northern) analysis using placental poly(A){sup +} RNA or total RNA from four different human fibroblast cell strains revealed a single 1.6-kb transcript. Genomic blots using DNA from each cell strain digested with either EcoRI or PstI revealed a complex pattern of cDNA-hydridizing restriction fragments. The genomic analysis for each enzyme was highly similar in all four human cell strains. In contrast, a single band was observed when genomic analysis was performed with the identical DNA digests with an actin gene probe. During cell proliferation there was an increase in the level of glycosylase mRNA that paralleled the increase in uracil DNA glycosylase enzyme activity. The isolation of the human uracil DNA glycosylase gene permits an examination of the structure, organization, and expression of a human DNA repair gene.
- OSTI ID:
- 6957930
- Journal Information:
- Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 86:22; ISSN 0027-8424; ISSN PNASA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
ANTIBODIES
AZINES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
CELL PROLIFERATION
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
DNA REPAIR
ENZYMES
FETAL MEMBRANES
GENE RECOMBINATION
GENE REGULATION
GENES
GLYCOSYL HYDROLASES
HETEROCYCLIC COMPOUNDS
HYBRIDIZATION
HYDROLASES
HYDROXY COMPOUNDS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MAN
MEMBRANES
MESSENGER-RNA
MOLECULAR BIOLOGY
MONOCLONAL ANTIBODIES
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLACENTA
PLASMIDS
PRIMATES
PURIFICATION
PYRIMIDINES
RADIOISOTOPES
RECOVERY
REPAIR
RNA
URACILS
VERTEBRATES