Chapter 17
Electroporation Transformation Protocol for Phaeodactylum
tricornutum
Hanhua Hu and Yufang Pan
Abstract
Genetic transformation system is very important for both basic biological research and commercial exploitation of diatoms. Here we describe a high-efficiency nuclear transformation method for the model diatom
Phaeodactylum tricornutum using an electroporation system, and the maximum transformation frequency
obtained is about 3 Â 10
À5 cells. The described protocol also provides some clue for developing electroporation transformation system in other eukaryotic microalgae.
Key words Transformation, Electroporation, Diatoms, Phaeodactylum tricornutum
1 Introduction
Diatoms are one of the most common types of phytoplankton. To
develop an efficient genetic transformation system is very important
for both basic biological research and commercial exploitation of
diatoms. At present, transformation of diatoms has been established for several diatom species, including Phaeodactylum tricornutum [1–4], Cylindrotheca fusiformis [5, 6], Thalassiosira
pseudonana [7], Chaetoceros sp. [8], Fistulifera sp. [9], Navicula
saprophila, and Cyclotella cryptica [10]. These established techniques for transporting DNA into diatom cells relied on microprojectile bombardment. However, microprojectile bombardment is
costly and not routinely available, and the transformation efficiency
is not high thus limiting its application in many laboratories.
Though high efficient electroporation-mediated transformation was achieved in eukaryotic Chlamydomonas reinhardtii
20 years ago [11], stable gene transfer by electroporation was
established in other eukaryotic microalgae only very recently,
including Nannochloropsis sp. [12–14] and some green algae. Niu
et al. [15] and Miyahara et al. [16] reported the transformation of
P. tricornutum by electroporation. However, the former did not
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_17,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
163
Electroporation Transformation Protocol for Phaeodactylum
tricornutum
Hanhua Hu and Yufang Pan
Abstract
Genetic transformation system is very important for both basic biological research and commercial exploitation of diatoms. Here we describe a high-efficiency nuclear transformation method for the model diatom
Phaeodactylum tricornutum using an electroporation system, and the maximum transformation frequency
obtained is about 3 Â 10
À5 cells. The described protocol also provides some clue for developing electroporation transformation system in other eukaryotic microalgae.
Key words Transformation, Electroporation, Diatoms, Phaeodactylum tricornutum
1 Introduction
Diatoms are one of the most common types of phytoplankton. To
develop an efficient genetic transformation system is very important
for both basic biological research and commercial exploitation of
diatoms. At present, transformation of diatoms has been established for several diatom species, including Phaeodactylum tricornutum [1–4], Cylindrotheca fusiformis [5, 6], Thalassiosira
pseudonana [7], Chaetoceros sp. [8], Fistulifera sp. [9], Navicula
saprophila, and Cyclotella cryptica [10]. These established techniques for transporting DNA into diatom cells relied on microprojectile bombardment. However, microprojectile bombardment is
costly and not routinely available, and the transformation efficiency
is not high thus limiting its application in many laboratories.
Though high efficient electroporation-mediated transformation was achieved in eukaryotic Chlamydomonas reinhardtii
20 years ago [11], stable gene transfer by electroporation was
established in other eukaryotic microalgae only very recently,
including Nannochloropsis sp. [12–14] and some green algae. Niu
et al. [15] and Miyahara et al. [16] reported the transformation of
P. tricornutum by electroporation. However, the former did not
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_17,
© Springer Science+Business Media, LLC, part of Springer Nature 2020
163