Chapter 20
Electroporation Procedures for Genetic Modification
of Green Algae (Chlorella spp.)
Liang Ji and Jianhua Fan
Abstract
The green algae of Chlorella spp. are usually very small (about 3–6 μm), typically have solid and thick cell
wall tissue; thus, neither the gene-gun method based on particle carrier nor the glass-bead transformation
method is suitable enough. Selecting the proper, effective strategy has always attracted researcher’s attention. Electroporation is currently the most widely used method for the transformation of algal species. The
principle of electroporation is that the cell membrane produces tiny holes by high-voltage pulses, which lead
to the introduction of exogenous DNA into cells. The method was proved by simple in principle and
effective in introducing foreign genes in several Chlorella species.
Key words Chlorella, Electrotransformation, Exogenous DNA, Plasmids, Genetic engineering,
Resistance selection
1 Introduction
Electrotransformation, through the action of a high-intensity electric field, instantaneously improves the permeability of the cell
membrane, thereby absorbing foreign molecules from the surrounding media.
This technique can transfer nucleotides, DNA and RNA, proteins, carbohydrates, dyes and virus particles into prokaryotic and
eukaryotic cells. Electrotransformation is a valuable and effective
alternative to other physical and chemical conversion methods,
which is widely used in the transformation of algal species. Neither
the gene-gun method nor the glass-bead transformation method is
suitable for Chlorella spp. [1, 2], but some of them have been
effectively transformed by electrotransformation like other algal
species [3–8]. Selectable markers and reporter genes are needed
to distinguish transformants; hence, the right choice of the resistance selection is also the key for successful transformation.
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_20,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
181
Electroporation Procedures for Genetic Modification
of Green Algae (Chlorella spp.)
Liang Ji and Jianhua Fan
Abstract
The green algae of Chlorella spp. are usually very small (about 3–6 μm), typically have solid and thick cell
wall tissue; thus, neither the gene-gun method based on particle carrier nor the glass-bead transformation
method is suitable enough. Selecting the proper, effective strategy has always attracted researcher’s attention. Electroporation is currently the most widely used method for the transformation of algal species. The
principle of electroporation is that the cell membrane produces tiny holes by high-voltage pulses, which lead
to the introduction of exogenous DNA into cells. The method was proved by simple in principle and
effective in introducing foreign genes in several Chlorella species.
Key words Chlorella, Electrotransformation, Exogenous DNA, Plasmids, Genetic engineering,
Resistance selection
1 Introduction
Electrotransformation, through the action of a high-intensity electric field, instantaneously improves the permeability of the cell
membrane, thereby absorbing foreign molecules from the surrounding media.
This technique can transfer nucleotides, DNA and RNA, proteins, carbohydrates, dyes and virus particles into prokaryotic and
eukaryotic cells. Electrotransformation is a valuable and effective
alternative to other physical and chemical conversion methods,
which is widely used in the transformation of algal species. Neither
the gene-gun method nor the glass-bead transformation method is
suitable for Chlorella spp. [1, 2], but some of them have been
effectively transformed by electrotransformation like other algal
species [3–8]. Selectable markers and reporter genes are needed
to distinguish transformants; hence, the right choice of the resistance selection is also the key for successful transformation.
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_20,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
181