TY - JOUR
T1 - Transformation of the rhizospheric Bacillus cereus sensu lato B25 strain using a room-temperature electrocompetent cells preparation protocol
AU - Morales-Ruiz, Estefanía
AU - López-Ceballos, Astrid
AU - Maldonado-Mendoza, Ignacio E.
N1 - Publisher Copyright:
© 2019 Elsevier Inc.
PY - 2019/9
Y1 - 2019/9
N2 - Bacterial transformation is a crucial step in the genetic manipulation of a bacterium. However, Gram-positive bacteria are difficult to transform and consequently many different methodologies have been developed. Here, we examined the transformation efficiencies of an electroporation protocol by varying three main factors: the composition of the electroporation buffer, the strength of the electric pulse, and the composition of the recovery media. Overall, transformation efficiency was enhanced when we prepared the electrocompetent cells at room temperature instead of an ice-cold temperature. The protocol detailed in this work was demonstrated to be applicable to another B. cereus strain and two other Bacillus species, and has the potential to be applied to other undomesticated Gram-positive and/or rhizospheric bacterial strains that are difficult to transform using current methodologies.
AB - Bacterial transformation is a crucial step in the genetic manipulation of a bacterium. However, Gram-positive bacteria are difficult to transform and consequently many different methodologies have been developed. Here, we examined the transformation efficiencies of an electroporation protocol by varying three main factors: the composition of the electroporation buffer, the strength of the electric pulse, and the composition of the recovery media. Overall, transformation efficiency was enhanced when we prepared the electrocompetent cells at room temperature instead of an ice-cold temperature. The protocol detailed in this work was demonstrated to be applicable to another B. cereus strain and two other Bacillus species, and has the potential to be applied to other undomesticated Gram-positive and/or rhizospheric bacterial strains that are difficult to transform using current methodologies.
KW - Bacillus
KW - Bacterial transformation
KW - Rhizosphere
UR - http://www.scopus.com/inward/record.url?scp=85070058835&partnerID=8YFLogxK
U2 - 10.1016/j.plasmid.2019.102435
DO - 10.1016/j.plasmid.2019.102435
M3 - Artículo
C2 - 31374235
AN - SCOPUS:85070058835
SN - 0147-619X
VL - 105
JO - Plasmid
JF - Plasmid
M1 - 102435
ER -