Nuclear Gene Transformation in the Dinoflagellate Oxyrrhis marina

Authors:
Brittany N Sprecher , Huan Zhang , Senjie Lin 
In:
Source:
Publication Date: ()
Issue: 8: 126
Research Area:
Basic Research
Platform:
4D-Nucleofector® X-Unit
Experiment

Electroporation was carried out using Lonza 4D-Nucleofector™ X Unit system in 16-well Nucleocuvette™ Strips using the manufacturer’s SG and Supplemental 1 solutions.

Taking all the data into consideration, the pulse codes that showed overall strongest performance were DS-134, DS-137, and DS-120. Nevertheless, we recommend the use of all seven settings (DS-137, DS-130, DS-138, DS-134, DS-150, ED-150, DS-120, no pulse control) in the first optimization tests for this species. For other algae to be studied, full optimization tests with the other available solutions should be utilized when using the 4D-Nucleofector™ X Unit.

Abstract

The lack of a robust gene transformation tool that allows proper expression of foreign genes and functional testing for the vast number of nuclear genes in dinoflagellates has greatly hampered our understanding of the fundamental biology in this ecologically important and evolutionarily unique lineage of microeukaryotes. Here, we report the development of a dinoflagellate expression vector containing various DNA elements from phylogenetically separate dinoflagellate lineages, an electroporation protocol, and successful expression of introduced genes in an early branching dinoflagellate, Oxyrrhis marina. This protocol, involving the use of Lonza's Nucleofector and a codon-optimized antibiotic resistance gene, has been successfully used to produce consistent results in several independent experiments for O. marina. It is anticipated that this protocol will be adaptable for other dinoflagellates and will allow characterization of many novel dinoflagellate genes.