Wendy Greenawalt

Data Blitz Presentation

"Accidental Discovery of a Plasmid that Replicates in Alteromonas macleodii"

Transformation of Altermonas macleodii by electroporation and heat shock have been anecdotally unsuccessful, so we attempted to create an in-frame deletion system based on conjugation and homologous recombination. Vector design was based on a known conjugal transfer vector. However, when successful conjugants were further analyzed, the vector had failed to integrate and proved to be self-replicating. We now have a shuttle vector that can be used for gain of function, heterologous expression, and complementation.

Poster Presentation

Cyclic-di-GMP is a widespread bacterial second messenger that regulates numerous cellular functions, including biofilm formation. However, little information exists on its ecological significance in marine microbes. Alteromonas macleodii serves as an ideal model organism to examine the role of c-di-GMP signaling in biofilm formation due to its particle-associated lifestyle. Attempts to generate in-frame deletion mutations through a conjugal mating system were not successful because the conjugal vector was unexpectedly able to replicate in A.macleodii. While this system has not successfully created knockout organisms, it does provide a vector that could be used for gain of function, heterologous expression, and complementation experiments.