A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers

Abstract Background The unicellular red alga Cyanidioschyzon merolae exhibits a very simple cellular and genomic architecture. In addition, procedures for genetic modifications, such as gene targeting by homologous recombination and inducible/repressible gene expression, have been developed. However...

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Autores principales:	Takayuki Fujiwara, Shunsuke Hirooka, Shin-ya Miyagishima
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Cotransformation Cyanidioschyzon merolae Genetic modification Photosynthetic eukaryote Unicellular red alga Botany QK1-989
Acceso en línea:	https://doaj.org/article/1bebee2ba0994c46b502895d13a404cb
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spelling	oai:doaj.org-article:1bebee2ba0994c46b502895d13a404cb2021-12-05T12:06:25ZA cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers10.1186/s12870-021-03365-z1471-2229https://doaj.org/article/1bebee2ba0994c46b502895d13a404cb2021-12-01T00:00:00Zhttps://doi.org/10.1186/s12870-021-03365-zhttps://doaj.org/toc/1471-2229Abstract Background The unicellular red alga Cyanidioschyzon merolae exhibits a very simple cellular and genomic architecture. In addition, procedures for genetic modifications, such as gene targeting by homologous recombination and inducible/repressible gene expression, have been developed. However, only two markers for selecting transformants, uracil synthase (URA) and chloramphenicol acetyltransferase (CAT), are available in this alga. Therefore, manipulation of two or more different chromosomal loci in the same strain in C. merolae is limited. Results This study developed a nuclear targeting and transformant selection system using an antibiotics blasticidin S (BS) and the BS deaminase (BSD) selectable marker by homologous recombination in C. merolae. In addition, this study has succeeded in simultaneously modifying two different chromosomal loci by a single-step cotransformation based on the combination of BSD and CAT selectable markers. A C. merolae strain that expresses mitochondrion-targeted mSCARLET (with the BSD marker) and mVENUS (with the CAT marker) from different chromosomal loci was generated with this procedure. Conclusions The newly developed BSD selectable marker enables an additional genetic modification to the already generated C. merolae transformants based on the URA or CAT system. Furthermore, the cotransformation system facilitates multiple genetic modifications. These methods and the simple nature of the C. merolae cellular and genomic architecture will facilitate studies on several phenomena common to photosynthetic eukaryotes.Takayuki FujiwaraShunsuke HirookaShin-ya MiyagishimaBMCarticleCotransformationCyanidioschyzon merolaeGenetic modificationPhotosynthetic eukaryoteUnicellular red algaBotanyQK1-989ENBMC Plant Biology, Vol 21, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Cotransformation Cyanidioschyzon merolae Genetic modification Photosynthetic eukaryote Unicellular red alga Botany QK1-989
spellingShingle	Cotransformation Cyanidioschyzon merolae Genetic modification Photosynthetic eukaryote Unicellular red alga Botany QK1-989 Takayuki Fujiwara Shunsuke Hirooka Shin-ya Miyagishima A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
description	Abstract Background The unicellular red alga Cyanidioschyzon merolae exhibits a very simple cellular and genomic architecture. In addition, procedures for genetic modifications, such as gene targeting by homologous recombination and inducible/repressible gene expression, have been developed. However, only two markers for selecting transformants, uracil synthase (URA) and chloramphenicol acetyltransferase (CAT), are available in this alga. Therefore, manipulation of two or more different chromosomal loci in the same strain in C. merolae is limited. Results This study developed a nuclear targeting and transformant selection system using an antibiotics blasticidin S (BS) and the BS deaminase (BSD) selectable marker by homologous recombination in C. merolae. In addition, this study has succeeded in simultaneously modifying two different chromosomal loci by a single-step cotransformation based on the combination of BSD and CAT selectable markers. A C. merolae strain that expresses mitochondrion-targeted mSCARLET (with the BSD marker) and mVENUS (with the CAT marker) from different chromosomal loci was generated with this procedure. Conclusions The newly developed BSD selectable marker enables an additional genetic modification to the already generated C. merolae transformants based on the URA or CAT system. Furthermore, the cotransformation system facilitates multiple genetic modifications. These methods and the simple nature of the C. merolae cellular and genomic architecture will facilitate studies on several phenomena common to photosynthetic eukaryotes.
format	article
author	Takayuki Fujiwara Shunsuke Hirooka Shin-ya Miyagishima
author_facet	Takayuki Fujiwara Shunsuke Hirooka Shin-ya Miyagishima
author_sort	Takayuki Fujiwara
title	A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
title_short	A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
title_full	A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
title_fullStr	A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
title_full_unstemmed	A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers
title_sort	cotransformation system of the unicellular red alga cyanidioschyzon merolae with blasticidin s deaminase and chloramphenicol acetyltransferase selectable markers
publisher	BMC
publishDate	2021
url	https://doaj.org/article/1bebee2ba0994c46b502895d13a404cb
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A cotransformation system of the unicellular red alga Cyanidioschyzon merolae with blasticidin S deaminase and chloramphenicol acetyltransferase selectable markers

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