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DEVELOPMENT OF BIOCHEMICAL AND IMMUNOLOGICAL ASSAYS FOR DNA RECOMBINATION AND REPAIR

Objective


Using affinity purified anti-recA antibodies, we identified a yeast protein which displayed several of the properties expected for a recombinase:
induction after ultraviolet irradiation;
increased expression in concomitance with recombination at meiosis;
nuclear localization.

The protein was partially purified but no activity associated with a recombinative function was detected. Analysis of the sequence of the corresponding gene indicated that the protein showing immunological cross reactivity to recA is the small subunit of ribonucleotide reductase.

Research conducted has been as follows:
purification and characterization of the eukaryotic analogues of recA protein;
development of immunological reagents and analysis of the induction of recA analogues following treatments which damage deoxyribonucleic acid (DNA);
cloning of the genes encoding recA protein analogues in yeast and mammalian cells;
development of a general method for the identification of proteins involved in DNA recombination and repair.
1. PURIFICATION AND CHARACTERIZATION OF THE EUKARYOTIC ANALOGS OF RECA PROTEIN.
2. DEVELOPMENT OF IMMUNOLOGICAL REAGENTS AND ANALYSIS OF THE INDUCTION OF THE RECA ANALOGS FOLLOWING DNA DAMAGING TREATMENTS.
3. CLONING OF THE GENES ENCODING THE ANALOGS OF RECA PROTEIN IN YEAST AND MAMMALIAN CELLS.
4. DEVELOPMENT OF A GENERAL METHOD FOR THE IDENTIFICATION OF PROTEINS INVOLVED IN DNA RECOMBINATION AND REPAIR.

EXPECTED BENEFITS

EVALUATION OF GENETIC RISKS FROM IONIZING RADIATION. DETERMINATION OF CELLULAR RADIOSENSITIVITY AND RELATION TO REPAIR.

Funding Scheme

CSC - Cost-sharing contracts

Coordinator

Universita Degli Studi Di Milano
Address
Via Festa Del Perdono 7
20122 Milano
Italy