Open Reading Frame Filtering Methods for Identification of Genic Sequences

Protein-Protein Interactions (PPIs) help understanding disease processes and their mechanisms. Ideally, researcher would like to understand the full network of PPIs that take place within a cell “interactome”, that is why Open Reading Frame (ORF) filtering method was utilized. ORF is a DNA fragment that lacks stop codon and has the potential to demonstrate a physiological interaction. ORFs are obtained by random shearing of DNA into small fragments. Fragments are filtered by insertion upstream of a selectable marker, to allow the survival of cells that only have an ORF. As vast majority of out of frame fragments encodes a premature stop codon. Here we present a method for filtering of genic ORFs ‘real gene' which results in a physiological protein. Furthermore, researchers thought fragments libraries rather than full-length libraries are perhaps counterintuitive, as expected to result in a high rate of false negatives. However, researchers have found fragments libraries rather than full-length reduce number of false negative interactions. Lastly, great advantage to note is while using fragmented library it allows localization of interaction site, offering a robust path for drug targets, treatments towards cancer and the arising resistance to antibiotics.