Establishment of a novel set of vectors for transformation of the dinoflagellateAmphidinium carterae

Abstract

Peridinin-containing dinoflagellate algae have a chloroplast genome formed from plasmid-like minicircles. This fragmented genome has allowed us to develop a genetic modification methodology involving the use of biolistics to introduce artificial minicircles inAmphidinium carterae(Nimmo et al., 2019). The previously reported artificial minicircles were based on native minicircles containing either thepsbAoratpBgene. Each artificial minicircle allowed expression of a single selectable marker instead ofpsbAoratpB. Here, we present two further artificial minicircles for use in transformation ofA. carterae. One is based on thepetDminicircle, allowing the expression of a single selectable marker. The second is based on the two-gene minicircle originally containingatpAandpetB, and allows the dual expression of a selectable marker and a gene of interest. Our research suggest that all of the 20 or so minicircles inA. carteraeare suitable for adaptation as artificial minicircles, allowing for the simultaneous introduction of multiple genes.