Objective 1) STUDY OF MOLECULAR INTERACTIONS OF ENVIRONMENTAL MUTAGENS AND CARCINOGENS OF DNA LEVEL. 2) TO RELATE BIOLOGICAL CONSEQUENCES OF CHEMICAL TO THE EXTENT AND SITE OF REACTION BETWEEN THE MUTAGENS AND DNA. The detection of deoxyribonucleic acid (DNA) alkylation and its mutagenic effects in defined sequences were investigated. Over 10 000 plaques were examined without a mutation being detected. After careful consideration of a number of possible explanations, it is now believed that, although the oligonucleotide used definitely contained a residue of O6meG, strand displacement may have occurred during in vitro replication. Because the destabilising effect of the lesion was more extensive than anticipated there was an effective loss of methylation at the stage of the double strand formation.1) DEVELOPMENT OF NEW ASSAY METHODOLOGY, SPECIFIC FOR 06-ALKYLGUANINETYPE DAMAGE BUT WITH RELATIVELY WIDE ALKYL-GROUP SPECIFICITY. THIS WOULD BE BASED ON THE USE OF THE ENZYME 06-ALKYLGUANINE-DNA ALKYLTRANSFERASE AND RADIOLABELLED SYNTHETIC OLIGONUCLEOTIDES AS PROBES FOR THIS TYPE OF LESION. 2) EXAMINATION, IN VITRO, OF THE BASE-CODING PROPERTIES (AND THE EFFECTS OF NEIGHBOURING NUCLEOTIDES ON THEM) OF IMPORTANT ALKYLATION LESIONS IN DEFINED SEQUENCES OF SYNTHETIC OLIGONUCLEOTIDES DURING DNA REPLICATION PAST THEM. 3) DETERMINATION AT THE MOLECULAR LEVEL OF THE MUTAGENIC EFFECTS OF ALKYLATION ON A SPECIFIC SEQUENCE, THROUGH IN VITRO ALKYLATION OF THE SEQUENCE, BACTERIAL TRANSFORMATION AND MUTANT SELECTION, DNA SEQUENCE ANALYSIS AND FURTHER ANALYSIS OF THE SEQUENCE-DEPENDENCE OF THE OBSERVED MUTAGENESIS A) BY MEASUREMENT OF ALKYLATION AT SPECIFIC SITES AND B) BY CONSTRUCTION OF SITE-SPECIFICALLY ALKYLATED VECTORS AND STUDY OF THEIR REPAIR AND MUTAGENIC PROPERTIES. Fields of science natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesgeneticsmutationnatural sciencesearth and related environmental sciencesenvironmental sciencespollutionnatural sciencesbiological sciencesgeneticsnucleotidesnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programme(s) FP1-ENVPROT 4C - Multiannual research and development programmes (EEC) in the field of the environment - Protection of the environment -, 1986-1990 Topic(s) Data not available Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator NATIONAL HELLENIC RESEARCH FOUNDATION EU contribution No data Address Vassileos Constantinou Avenue 48 ATHENS Greece See on map Links Website Opens in new window Total cost No data