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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Jeon S, Kang NK, Suh WI et al (2019) Optimization of electroporation-based multiple pulses and further improvement of transformation efficiency using bacterial conditioned medium for Nannochloropsis salina. J Appl Phycol 31:1153–1161
Fromm ME, Taylor LP, Walbot V (1986) Stable transformation of maize after gene transfer by electroporation. Nature 319:791–793
Kotnik T, Frey W, Sack M et al (2015) Electroporation-based applications in biotechnology. Trends Biotechnol 33:480–488
Xue J, Balamurugan S, Li DW et al (2017) Glucose-6-phosphate dehydrogenase as a target for highly efficient fatty acid biosynthesis in microalgae by enhancing NADPH supply. Metab Eng 41:212–221
Yoon K, Han D, Li Y et al (2012) Phospholipid: diacylglycerol acyltransferase is a multifunctional enzyme involved in membrane lipid turnover and degradation while synthesizing triacylglycerol in the unicellular green microalga Chlamydomonas reinhardtii. Plant Cell 24:3708–3724
Li DW, Cen SY, Liu YH et al (2016) A type 2 diacylglycerol acyltransferase accelerates the triacylglycerol biosynthesis in heterokont oleaginous microalga Nannochloropsis oceanica. J Biotechnol 229:65–71
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Li, DW., Balamurugan, S., Zheng, JW., Yang, WD., Liu, JS., Li, HY. (2020). Rapid and Effective Electroporation Protocol for Nannochloropsis oceanica. In: Li, S., Chang, L., Teissie, J. (eds) Electroporation Protocols. Methods in Molecular Biology, vol 2050. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9740-4_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9740-4_19
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9739-8
Online ISBN: 978-1-4939-9740-4
eBook Packages: Springer Protocols