Development of a 3’3’‐Cyclic GMP‐AMP Enzyme Linked Immunoassay Reveals Phage Infection Reduces DncV Activity

Abstract East Lansing, MI 48824 Cyclic di‐nucleotides (CDNs) are central signaling molecules in organisms spanning the tree of life. In bacteria, CDNs mediate many important physiological functions such as biofilm formation, motility, and virulence. also implicated activation cellular biological defense systems both bacteria eukaryotes. CDN 3’3’‐cyclic GMP‐AMP (3’3’‐cGAMP) activates a putative phage system Vibrio cholerae controls processes Geobacter sulfurreducens Bdellovibrio bacteriovorus . Therefore, access to rapid, accurate 3’3’‐cGAMP quantification assay would enable further studies this molecule. Here, we describe validation novel enzyme linked immunoassay (ELISA) developed by Cayman Chemicals. We demonstrate that concentrations determined ELISA strongly correlate with those obtained using liquid chromatography‐tandem mass spectrometry (LC‐MS/MS). Furthermore, during these show V. synthase, DncV, when expressed itself Escherichia coli , is not activated infection.