Electro-transformation of Lactobacillus spp.

Overview

While there is a wide variety of methods used for the electrotransformation of Lactobacillus spp., there are a few steps all of them share. This page will list many of these protocols in a standard format so that you can compare them and choose the one that works for you. In some cases an apples to apples comparison may not be acceptable (e.g. the differences in electroporators can be dramatic) so be sure to check the actual paper.

Procedures

Josson et al. (1989)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Author (Year)

• Growth -
• Wash -
• Buffer -
• Pulse -
• Recovery -
• Reference -

Notes

All questions, input and feedback are welcome!

1. Centrifugation at 4000 rpm for 2 minutes was sufficient to pellet the competent cells to give a clear supernatant.

References

This protocol is based mostly on the protocols described by Thompson and Collins (1996) and Mason et al. (2004) but other literature are also listed.

Alegre et al (FEMS Microbiology Letters 241 (2004) 73-77)

Aukrust, T. and H. Blom (1992). "TRANSFORMATION OF LACTOBACILLUS STRAINS USED IN MEAT AND VEGETABLE FERMENTATIONS." Food Research International 25(4): 253-261.

Berthier, F., M. Zagorec, et al. (1996). "Efficient transformation of Lactobacillus sake by electroporation." Microbiology-Uk 142: 1273-1279.

Josson, K., T. Scheirlinck, et al. (1989). "CHARACTERIZATION OF A GRAM-POSITIVE BROAD-HOST-RANGE PLASMID ISOLATED FROM LACTOBACILLUS-HILGARDII." Plasmid 21(1): 9-20.

Mason, C. K., M. A. Collins, et al. (2005). "Modified electroporation protocol for Lactobacilli isolated frorn the chicken crop facilitates transformation and the use of a genetic tool." Journal of Microbiological Methods 60(3): 353-363.

Posno, M., R. J. Leer, et al. (1991). "INCOMPATIBILITY OF LACTOBACILLUS VECTORS WITH REPLICONS DERIVED FROM SMALL CRYPTIC LACTOBACILLUS PLASMIDS AND SEGREGATIONAL INSTABILITY OF THE INTRODUCED VECTORS." Applied and Environmental Microbiology 57(6): 1822-1828.

Thompson, K. and M. A. Collins (1996). "Improvement in electroporation efficiency for Lactobacillus plantarum by the inclusion of high concentrations of glycine in the growth medium." Journal of Microbiological Methods 26(1-2): 73-79.

Wei, M. Q., C. M. Rush, et al. (1995). "AN IMPROVED METHOD FOR THE TRANSFORMATION OF LACTOBACILLUS STRAINS USING ELECTROPORATION." Journal of Microbiological Methods 21(1): 97-109.

Contact

• morto077@uottawa.ca

• mas853@psu.edu

or instead, discuss this protocol. -->