3. Cells are thoroughly washed with 375 mM sorbitol prior to
electroporation to remove the influence of salts which could
affect the electroporation.
4. The volume of the desired DNA to be electroporated should
not exceed 10% of the total volume, which in turn affect the
electroporation delivery.
5. After electroporation, the cells are incubated in dark and cells
are gently rotated to avoid damage.
6. After the subculture of the electroporated cells into solid selection medium, the surviving colonies with uniform phenotypes
are further assessed by molecular analysis and the cells are
subjected to further analyses.
References
1. 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
2. Fromm ME, Taylor LP, Walbot V (1986) Stable
transformation of maize after gene transfer by
electroporation. Nature 319:791–793
3. Kotnik T, Frey W, Sack M et al (2015)
Electroporation-based applications in biotechnology. Trends Biotechnol 33:480–488
4. 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
5. 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
6. 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
Electroporation of Nannochloropsis
179
electroporation to remove the influence of salts which could
affect the electroporation.
4. The volume of the desired DNA to be electroporated should
not exceed 10% of the total volume, which in turn affect the
electroporation delivery.
5. After electroporation, the cells are incubated in dark and cells
are gently rotated to avoid damage.
6. After the subculture of the electroporated cells into solid selection medium, the surviving colonies with uniform phenotypes
are further assessed by molecular analysis and the cells are
subjected to further analyses.
References
1. 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
2. Fromm ME, Taylor LP, Walbot V (1986) Stable
transformation of maize after gene transfer by
electroporation. Nature 319:791–793
3. Kotnik T, Frey W, Sack M et al (2015)
Electroporation-based applications in biotechnology. Trends Biotechnol 33:480–488
4. 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
5. 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
6. 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
Electroporation of Nannochloropsis
179