Parámetros de electroporación para especies de microalgas marinas, usando como modelo Porphyridium cruentum
Date
2023-11-20
Authors
Palacio Quintero, Sofía
Obando Montoya, Erika Juliana
Ortiz Mejía, Diego Alejandro
Journal Title
Journal ISSN
Volume Title
Publisher
Universidad CES
Abstract
Genetic transformation of cells is a technique that has allowed the development of processes
to produce heterologous proteins with functional interest and in an efficient manner. One of
the methods that facilitates this process is electroporation, a common and efficient technique.
It employs electric field pulses (PEF), which generate a potential change that alters the lipid
bilayer, thus creating pores that allow the temporary entry of macromolecules. Whether or
not electroporation occurs is linked to the combination of the characteristics of the electric
field (number, duration, time between pulses, etc.), and the characteristics of the cellular state
(geometry, temperature, etc.). Permeabilization can be reversible or irreversible, and
reversible permeabilization is of particular relevance for successful transformation, since in
this state the cells open their pores and subsequently their membranes regenerate, avoiding
cell lysis. In the search for organisms that present interesting characteristics and on which the
different transformation methodologies can be applied, we find the red microalgae
Porphyridium cruentum. It has been highly studied for the functionality of its metabolites, its
high growth rates, which can be regulated, and its resistance to high salinity media. Therefore,
the objective of this study was to determine the effect of the electric field and the variation
of electroporation parameters on the permeabilization and viability of the cells of this
microalga. For this purpose, the cells were subjected to four treatments: T1 and T2 that were
not electroporated, and T3 and T4 that were electroporated (2kV/cm; 10 and 50 pulses;
duration of 30 and 150 µs). After the treatments, the cells were monitored. In addition, it was
possible to design a PEF-generating equipment and a base protocol for future electroporation
studies in cells of these microalgae and similar organisms.
Description
Keywords
Electroporation, Porphyridium cruentum, Electric field, Microalgae, Genetic transformation, Protocol