Life Sciences Commons^™

Cloning, Expression, And Purification Of Fadk And Its Application In Coa-Rna Capture, Jon-Michael L. Stork

Honors Theses

Coenzyme A (CoA) is an important enzyme cofactor involved in acyl transfer reactions. Recently, CoA and its various thioesters were found to exist at the 5’ end of RNA molecules. Although the function of these molecules is still unknown due to difficulties in their isolation, their existence at the 5’ RNA end reveals potentially novel biological roles of RNA. We are exploiting the broad substrate tolerance of the medium chain fatty acid-coenzyme A ligase (FadK), hypothesizing that this enzyme will accept biotinylated medium chain fatty acid substrates, creating biotin-tagged CoA-RNA. This modified CoA-RNA can later be captured using streptavidin affinity …