Cloning of Drosophila Gene which Code for UV-damaged DNA Binding Protein.

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Using a gel electrophoresis DNA band shift assay, we have identified two DNA binding protein complex in Drosophila embryonic cell which have a high affinity for UV-irradiated, double-stranded DNA. Screening Drosophila mutants which are deficient in DNA repair has identified five mutants which lack either of the two protein complex. Four excision repair deficient mutants (mus201, phr, mus308 and mus205) lack one protein complex (Factor 2). Another protein complex (Factor 1) was not detectable in post-replication repair deficient mutant mus104. These findings suggest a possible involvement of t he gene product in lesion recoghition and DNA repair of UV-light induced photoproduct.Using heparin agarose column chromatography, we can separate each factor into different fraction. The fraction which contain Factor 2, that is the fraction which have ability to form band 2, was further purified by UV-irradiated DNA affinity column chromatography. In the fraction which has ability to form band 2, we can identify only one protein band in silver stained SDS-PAGE gel. The molecular mass was assummed to be between 45k and 50k dalton. We assumed this protein is Factor 2.