Chapter 19
Rapid and Effective Electroporation Protocol
for Nannochloropsis oceanica
Da-Wei Li, Srinivasan Balamurugan, Jian-Wei Zheng, Wei-Dong Yang,
Jie-Sheng Liu, and Hong-Ye Li
Abstract
Electroporation refers to the application of high strength electric pulse to create transient pores in the
membrane, thereby enabling the passage of hydrophilic molecules into the cells. Based on the properties of
cell and cell wall, the electroporation parameters vary among the algal species. Here, we demonstrated the
optimized protocol for successful introduction of recombinant DNA (~5000 bp) into Nannochloropsis
oceanica. The linearized recombinant plasmid that harbors eGFP and Bh-sle as the reporter and marker
gene, respectively, was electroporated into the electrocompetent N. oceanica cells at voltage of 2200 V,
50 μF, resistance at 600 Ω using electroporator, and the transformed cells were then screened by molecular
analysis. The report exemplifies a straightforward and reliable electroporation strategy for generating
transgenic N. oceanica cells.
Key words DNA delivery, Algal cell transformation, Electroporation, Oleaginous microalgae,
Nannochloropsis
1 Introduction
Development of feasible DNA delivery strategy is pivotal for
exploiting the commercial potential of microalgae. Among the
commonly used techniques such as Agrobacterium-mediated transformation and particle bombardment, electroporation has been
considered an effective tool for algal transformation [1]. Electroporation involves the application of strong electric pulse to transiently
increase the electric conductivity and permeability of cellular membrane, thereby permitting the diffusion of DNA molecules into the
cells [2]. Even though the transformation efficiency of electroporation in microalgae was found to be lower than prokaryotic models,
the protocol was found to be adequate for the generation of transgenic lines [3]. Amidst, Chlamydomonas reinhardtii and Phaeodactylum tricornutum have been employed as the model microalgal
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_19,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
175
Rapid and Effective Electroporation Protocol
for Nannochloropsis oceanica
Da-Wei Li, Srinivasan Balamurugan, Jian-Wei Zheng, Wei-Dong Yang,
Jie-Sheng Liu, and Hong-Ye Li
Abstract
Electroporation refers to the application of high strength electric pulse to create transient pores in the
membrane, thereby enabling the passage of hydrophilic molecules into the cells. Based on the properties of
cell and cell wall, the electroporation parameters vary among the algal species. Here, we demonstrated the
optimized protocol for successful introduction of recombinant DNA (~5000 bp) into Nannochloropsis
oceanica. The linearized recombinant plasmid that harbors eGFP and Bh-sle as the reporter and marker
gene, respectively, was electroporated into the electrocompetent N. oceanica cells at voltage of 2200 V,
50 μF, resistance at 600 Ω using electroporator, and the transformed cells were then screened by molecular
analysis. The report exemplifies a straightforward and reliable electroporation strategy for generating
transgenic N. oceanica cells.
Key words DNA delivery, Algal cell transformation, Electroporation, Oleaginous microalgae,
Nannochloropsis
1 Introduction
Development of feasible DNA delivery strategy is pivotal for
exploiting the commercial potential of microalgae. Among the
commonly used techniques such as Agrobacterium-mediated transformation and particle bombardment, electroporation has been
considered an effective tool for algal transformation [1]. Electroporation involves the application of strong electric pulse to transiently
increase the electric conductivity and permeability of cellular membrane, thereby permitting the diffusion of DNA molecules into the
cells [2]. Even though the transformation efficiency of electroporation in microalgae was found to be lower than prokaryotic models,
the protocol was found to be adequate for the generation of transgenic lines [3]. Amidst, Chlamydomonas reinhardtii and Phaeodactylum tricornutum have been employed as the model microalgal
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_19,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
175