Abstract
Electroporation is a highly efficient delivery method for transformation in various cell types; however, in microalgae there is lack of optimized electroporation parameters due to cell wall, protoplast preparation and viability. Therefore, we optimized electroporation conditions for transforming microalgae using Chlamydomonas reinhardtii strains of wild type and mutant (cell wall deficient). To investigate the effects of molecule size, calcein (623 Da) and fluorescein isothiocyanate-dextran (FITC-dextran, 40 kDa) were used and various electroporation parameters were applied such as different voltage and pulse length and molecule uptake pattern and cell viability were observed. Cell wall is insignificant in case of small sized molecule uptake as noticed by 1.25 kV/cm and 30 ms for both strains, whereas for larger molecules by 1.5 and 2 kV/cm and 30 ms for mutant and wild type, respectively. In terms of viability, there was no significant difference in both the strains on applied electroporation parameters. The controlled parameters corresponding to 1.5 to 2.0 kV/cm and 20 to 30 ms could be used to deliver macromolecules (DNA, proteins) into cells effectively.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
O.R. Inderwildi and D. A. King, Energy Environ. Sci., 2, 343 (2009).
J. Lü, C. Sheahan and P. Fu, Energy Environ. Sci., 4, 2451 (2011).
T. M. Mata, A. A. Martins and N. S. Caetano, Renewable and Sustainable Energy Reviews, 14, 217 (2010).
Y. Chisti, Biotechnol. Adv., 25, 294 (2007).
L. L. Beer, E. S. Boyd, J.W. Peters and M.C. Posewitz, Curr. Opin. Biotechnol., 20, 264 (2009).
S.W. D. S. C. Davis and R.G. Boundy, Ed., Transportation energy data book, the Center for Transportation Analysis of the Oak Ridge National Laboratory (2012).
A. Demirbas, Energy Policy, 35, 4661 (2007).
G. Knothe, C. A. Sharp and T.W. Ryan, Energy Fuels, 20, 403 (2006).
R. Radakovits, R. E. Jinkerson, A. Darzins and M. C. Posewitz, Eukaryot Cell., 9, 486 (2010).
S. Qin, H. Lin and P. Jiang, Biotechnol. Adv., 30, 1602 (2012).
E.H. Harris, Annual Review of Plant Physiology and Plant Molecular Biology, 52, 363 (2001).
J. K. Hoober, Science, 246, 1503 (1989).
J. Shrager, C. Hauser, C.W. Chang, E. H. Harris, J. Davies, J. McDermott, R. Tamse, Z. Zhang and A. R. Grossman, Plant Physiol., 131, 401 (2003).
A. R. Grossman, M. Croft, V. N. Gladyshev, S. S. Merchant, M. C. Posewitz, S. Prochnik and M. H. Spalding, Curr. Opin. Plant Biol., 10, 190 (2007).
J. D. Rochaix and J. Vandillewijn, Nature, 296, 70 (1982).
E. Neumann, M. Schaeferridder, Y. Wang and P. H. Hofschneider, Embo. J., 1, 841 (1982).
U. Zimmermann, G. Pilwat and F. Riemann, Biochimica Et Biophysica Acta, 375, 209 (1975).
J. Gehl, Acta Physiologica Scandinavica, 177, 437 (2003).
L. E. Brown, S. L. Sprecher and L.R. Keller, Molecular and Cellular Biology, 11, 2328 (1991).
K. Shimogawara, S. Fujiwara, A. Grossman and H. Usuda, Genetics, 148, 1821 (1998).
P. J. Canatella, J. F. Karr, J. A. Petros and M.R. Prausnitz, Biophysical J., 80, 755 (2001).
H. R. Azencott, G. F. Peter and M. R. Prausnitz, Ultrasound in Medicine and Biology, 33, 1805 (2007).
C. Yang, H. A. Owen and P. Yang, J. Cell. Biol., 180, 403 (2008).
A. Lelong, H. Hegaret and P. Soudant, Research in Microbiology, 162, 969 (2011).
Z. Kolber and P.G. Falkowski, Limnology and Oceanography, 38, 1646 (1993).
J. P. Richard, K. Melikov, E. Vives, C. Ramos, B. Verbeure, M. J. Gait, L.V. Chernomordik and B. Lebleu, J. Biological Chem., 278, 585 (2003).
Y. Sun, Z.Y. Yang, X. S. Gao, Q.Y. Li, Q. Q. Zhang and Z. K. Xu, Molecular Biotechnology, 30, 185 (2005).
O. Kilian, C. S. E. Benemann, K. K. Niyogi and B. Vick, Proceedings of the National Academy of Sciences of the United States of America, 108, 21265 (2011).
J. C. Weaver and Y. A. Chizmadzhev, Bioelectrochemistry and Bioenergetics, 41, 135 (1996).
B. C. Monk, W. S. Adair, R. A. Cohen and U.W. Goodenough, Planta, 158, 517 (1983).
Author information
Authors and Affiliations
Corresponding author
Additional information
This paper is dedicated to commemorate Prof. Ji-Won Yang (KAIST) for his retirement.
Rights and permissions
About this article
Cite this article
Jeon, K., Suresh, A. & Kim, YC. Highly efficient molecular delivery into Chlamydomonas reinhardtii by electroporation. Korean J. Chem. Eng. 30, 1626–1630 (2013). https://doi.org/10.1007/s11814-013-0098-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11814-013-0098-4