Developing an efficient electroporation protocol for a model diatom algae.
Category: Research Poster
Author(s): Brooke Sturtz, Dane Ferguson, Matthew Herbert
Presenter(s): Brooke Sturtz, Dane Ferguson, Matthew Herbert
Mentors(s): Graham Peers, Andrew Paton
Diatoms like Phaeodactylum tricornutum utilize a brown light-harvesting carotenoid pigment called fucoxanthin to best absorb wavelengths of blue-green light in the ocean. Knockouts of the Phaeodactylum gene vdl2 lose the ability to produce fucoxanthin and are bright green in color; one specific strain of the vdl2 mutant has spontaneously reverted back to the brown color and has restored synthesis of fucoxanthin (named the fuc1 suppressor mutation). Our lab has begun using a newly published electroporation protocol to identify the gene associated with this mutation via expressing cDNA libraries from the fuc1 strain into a vdl2 background. We will report our efforts into improving the efficiency of this protocol. This includes testing the transformation efficiency with cells with grown in liquid or on plates, varying amounts of cDNA, and changing recovery tube sizes. This will aid in the development of tools used in other diatom-related molecular biology projects.