Virus-inspired transfection of a marine alga: isolation of EhV-207 viral envelope

Abstract

Emiliana huxleyi, a marine coccolithophore native to nearly all the world’s oceans, plays a key role in carbon fixation. Coccolithoviruses, such as EhV-207, infect and terminate E. huxleyi blooms. Our goal is to genetically modify E. huxleyi cells through a DNA delivery system characteristic of viruses like EhV-207. In this project, we attempt to isolate and visualize the lipid envelope of the EhV- 207 virion, which can, in turn, provide a suitable carrier for plasmid DNA to transfect E. huxleyi cells. Initially, very few virus particles were observed under the microscope, much fewer than anticipated. We describe here the process for cultivating, purifying, staining, and mounting the virus particles for microscopy and our efforts to troubleshoot the low yield of visible virus particles.