Chapter 16
Transformation of the Model Microalga Chlamydomonas
reinhardtii Without Cell-Wall Removal
Takashi Yamano and Hideya Fukuzawa
Abstract
The green alga Chlamydomonas reinhardtii has been widely used to study many biological processes,
including photosynthesis, flagellar motility, sexual reproduction, metabolism, and genetics. Here, we
describe a step-by-step protocol of rapid and efficient transformation method for wild type cell-walled
Chlamydomonas strains without cell-wall removal using a square electric pulses-generating electroporator.
This method could be applied to the transformation of other industrially useful algae including diatom by
optimizing the electric conditions.
Key words Chlamydomonas reinhardtii, Electroporation, Photosynthesis, Protist, Square electric
pulse
1 Introduction
With the growing emphasis on biofuel production by algae, it is
increasingly important to transform algae rapidly and efficiently. In
particular, the single cell green alga Chlamydomonas reinhardtii
(hereafter Chlamydomonas) is widely used to study the fundamental
biological processes of photosynthesis, cell cycle, organelle function/interaction, and cell motility as well as of biofuel production.
So far, genome sequences of the nucleus, chloroplast, and mitochondrion in Chlamydomonas are available [1] and three methods
of DNA-mediated transformation have been reported: bombardment with DNA-coated microprojectiles [2, 3], vortexing with
glass beads [4], and electroporation [5]. For nuclear genome transformation, the transformation efficiencies of the glass beads and
electroporation are approximately 10
3 and 10
5 transformants per
μg DNA, respectively [5]. Although electroporation-based transformation has been widely used, these procedures require the use of
cell-wall-less (cw) mutant strains or the removal of cell wall from
wild-type cells by treatment with the zinc-containing
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_16,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
155
Transformation of the Model Microalga Chlamydomonas
reinhardtii Without Cell-Wall Removal
Takashi Yamano and Hideya Fukuzawa
Abstract
The green alga Chlamydomonas reinhardtii has been widely used to study many biological processes,
including photosynthesis, flagellar motility, sexual reproduction, metabolism, and genetics. Here, we
describe a step-by-step protocol of rapid and efficient transformation method for wild type cell-walled
Chlamydomonas strains without cell-wall removal using a square electric pulses-generating electroporator.
This method could be applied to the transformation of other industrially useful algae including diatom by
optimizing the electric conditions.
Key words Chlamydomonas reinhardtii, Electroporation, Photosynthesis, Protist, Square electric
pulse
1 Introduction
With the growing emphasis on biofuel production by algae, it is
increasingly important to transform algae rapidly and efficiently. In
particular, the single cell green alga Chlamydomonas reinhardtii
(hereafter Chlamydomonas) is widely used to study the fundamental
biological processes of photosynthesis, cell cycle, organelle function/interaction, and cell motility as well as of biofuel production.
So far, genome sequences of the nucleus, chloroplast, and mitochondrion in Chlamydomonas are available [1] and three methods
of DNA-mediated transformation have been reported: bombardment with DNA-coated microprojectiles [2, 3], vortexing with
glass beads [4], and electroporation [5]. For nuclear genome transformation, the transformation efficiencies of the glass beads and
electroporation are approximately 10
3 and 10
5 transformants per
μg DNA, respectively [5]. Although electroporation-based transformation has been widely used, these procedures require the use of
cell-wall-less (cw) mutant strains or the removal of cell wall from
wild-type cells by treatment with the zinc-containing
Shulin Li et al. (eds.), Electroporation Protocols: Microorganism, Mammalian System, and Nanodevice,
Methods in Molecular Biology, vol. 2050, https://doi.org/10.1007/978-1-4939-9740-4_16,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
155