Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i>
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 linea...
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doaj-d4b3236af82a407493b5051401f6ab5d2020-11-25T02:06:05ZengMDPI AGMicroorganisms2076-26072020-01-018112610.3390/microorganisms8010126microorganisms8010126Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i>Brittany N. Sprecher0Huan Zhang1Senjie Lin2Department of Marine Sciences, University of Connecticut, 1080 Shennecossett Rd, Groton, CT 06340, USADepartment of Marine Sciences, University of Connecticut, 1080 Shennecossett Rd, Groton, CT 06340, USADepartment of Marine Sciences, University of Connecticut, 1080 Shennecossett Rd, Groton, CT 06340, USAThe 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, <i>Oxyrrhis marina</i>. 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 <i>O. marina</i>. It is anticipated that this protocol will be adaptable for other dinoflagellates and will allow characterization of many novel dinoflagellate genes.https://www.mdpi.com/2076-2607/8/1/126dinoflagellatesgene transformationmolecular biologymarine biology |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Brittany N. Sprecher Huan Zhang Senjie Lin |
spellingShingle |
Brittany N. Sprecher Huan Zhang Senjie Lin Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> Microorganisms dinoflagellates gene transformation molecular biology marine biology |
author_facet |
Brittany N. Sprecher Huan Zhang Senjie Lin |
author_sort |
Brittany N. Sprecher |
title |
Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> |
title_short |
Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> |
title_full |
Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> |
title_fullStr |
Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> |
title_full_unstemmed |
Nuclear Gene Transformation in the Dinoflagellate <i>Oxyrrhis marina</i> |
title_sort |
nuclear gene transformation in the dinoflagellate <i>oxyrrhis marina</i> |
publisher |
MDPI AG |
series |
Microorganisms |
issn |
2076-2607 |
publishDate |
2020-01-01 |
description |
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, <i>Oxyrrhis marina</i>. 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 <i>O. marina</i>. It is anticipated that this protocol will be adaptable for other dinoflagellates and will allow characterization of many novel dinoflagellate genes. |
topic |
dinoflagellates gene transformation molecular biology marine biology |
url |
https://www.mdpi.com/2076-2607/8/1/126 |
work_keys_str_mv |
AT brittanynsprecher nucleargenetransformationinthedinoflagellateioxyrrhismarinai AT huanzhang nucleargenetransformationinthedinoflagellateioxyrrhismarinai AT senjielin nucleargenetransformationinthedinoflagellateioxyrrhismarinai |
_version_ |
1724935132628910080 |