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), Vol. 86:22; ISSN 0027-8424
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA REPAIR
MOLECULAR BIOLOGY
GENES
PURIFICATION
GLYCOSYL HYDROLASES
GENE REGULATION
CELL PROLIFERATION
DNA HYBRIDIZATION
GENE RECOMBINATION
MAN
MESSENGER-RNA
MONOCLONAL ANTIBODIES
PHOSPHORUS 32
PLACENTA
PLASMIDS
URACILS
ANIMALS
ANTIBODIES
AZINES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
ENZYMES
FETAL MEMBRANES
HETEROCYCLIC COMPOUNDS
HYBRIDIZATION
HYDROLASES
HYDROXY COMPOUNDS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MEMBRANES
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PHOSPHORUS ISOTOPES
PRIMATES
PYRIMIDINES
RADIOISOTOPES
RECOVERY
REPAIR
RNA
VERTEBRATES
550201* - Biochemistry- Tracer Techniques