%0 Journal Article %T Construction of Vector for Functional Analysis of the Intrinsically Bent DNA in the ACE3 Replicator from <i>Drosophila melanogaster</i> %A Douglas Vin¨ªcius Bassalobre de Freitas %A Fabr¨ªcia Gimenes %A Adriana Fiorini %A Fabiana dos Santos Rando %A Quirino Alves de Lima Neto %A Maria Aparecida Fernandez %J Open Journal of Genetics %P 8-15 %@ 2162-4461 %D 2014 %I Scientific Research Publishing %R 10.4236/ojgen.2014.41002 %X
DNA replication is a crucial process for species survival, nevertheless it is not clear which factors define origin selection in multicellular eukaryotes. Developmental gene amplification systems, such as the one described during ovarian follicles development in Drosophila melanogaster, are useful tools for studying of DNA replication process in these organisms. We previously described that the well characterized third chromosome amplified domain of D. melanogater displays three intrinsically bent DNA sites: b1, localized at an amplification control element (ACE3), b2 and b3, both localized at the preferential origin ori-¦Â. This proposal aimed to construct a Drosophila transformation vector, which contains a short deletion at the ACE3, in order to reduce the intrinsically bent DNA site b1, and analyze the functional role of this site in the gene amplification process. Through a series of cloning steps, we obtained a Big Parent vector derivative, containing a deletion at the positions 176-