Antibody to a human DNA repair protein allows for cloning of a Drosophila cDNA that encodes an apurinic endonuclease
- Dept. of Biochemistry and Biophysics, Loyola Univ. Medical School, Maywood, IL (US)
- Louisiana State Univ., Baton Rouge, LA (USA). Dept. of Biochemistry
The cDNA of a Drosophila DNA repair gene, AP3, was cloned by screening an embryonic lambda gt11 expression library with an antibody that was originally prepared against a purified human apurinicapyrimidine (AP) endonuclease. The 1.2-kilobase (kb) AP3 cDNA mapped to a region on the third chromosome where a number of mutagen-sensitive alleles were located. The cDNA clone yielded an in vitro translation product of 35,000 daltons, in agreement with the predicted size of the translation product of the only open reading frame of AP3, and identical to the molecular size of an AP endonuclease activity recovered following sodium dodecyl sulfate-polyacrymalide gel electrophoresis of Drosophilia extracts. The C-terminal portion of the predicted protein contained regions of presumptive DNA-binding domains, while the DNA sequence at the amino end of AP3 showed similarity to the Escherichia coli recA gene. AP3 is expressed as an abundant 1.3-kb mRNA that is detected throughout the life cycle of Drosophila melanogaster. Another 3.5-klb mRNA also hybridized to the AP3 cDNA, but species was restricted to the early stages of development.
- OSTI ID:
- 5563861
- Journal Information:
- Molecular and Cellular Biology; (USA), Vol. 9:3; ISSN 0270-7306
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DROSOPHILA
DNA-CLONING
ENDONUCLEASES
GENETIC MAPPING
GENE RECOMBINATION PROTEINS
ANTIBODIES
BIOSYNTHESIS
ENZYME ACTIVITY
ESCHERICHIA COLI
GENES
MESSENGER-RNA
ANIMALS
ARTHROPODS
BACTERIA
CLONING
DIPTERA
DNA HYBRIDIZATION
DNA-ASE
ENZYMES
ESTERASES
FLIES
FRUIT FLIES
HYBRIDIZATION
HYDROLASES
INSECTS
INVERTEBRATES
MAPPING
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PROTEINS
RNA
SYNTHESIS
550400* - Genetics
550200 - Biochemistry