Optimization and comparison of two electrotransformation methods for lactobacilli

Heravi, R. M. and Nasiraii, L. R. and Sankian, M. and Kermanshahi, H. and Varasteh, A. R. (2012) Optimization and comparison of two electrotransformation methods for lactobacilli. Biotechnology, 11 (1). pp. 50-54.

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Abstract

To date, lactobacilli are widely used in food industries and new probiotic products; hence these are considered as an attractive target for genetic modifications. This study was conducted to improve electroporation of probiotic lactobacilli which is a necessary prerequisite for genetic manipulations. Four strains of probiotic lactobacilli from different sources were grown in Man Rogosa and Sharp (MRS) broth medium containing glycine, as a cell wall weakening agent or a pulse of glycine for evaluation of glycine effect on electroporation efficiency. After evaluation of various parameters such as washing buffers, washing times, electric field strength, pulse duration and plasmid concentration, a practical electroporation protocol was presented to improve electrotransformation of lactobacilli in comparison with two standard protocols. This new protocol exhibited higher transformation efficiency (transformants/μg plasmids) than standard methods (p<0.05) with no differences between species (Lb. casei, Lb. crispatus, Lb. salivarus and L. rhamnosus). The pulse of glycine had no effect on the number of the transformants in three methods. In contrast to plasmid concentration, plasmid size had no influence on the transformation efficiency. The modified method enabled to transform plasmid into the resistant lactobacilli against transformation. These bacteria have potential for bioengineering research to improve special performance. © 2012 Asian Network for Scientific Information.

Item Type: Article
Additional Information: Cited By :5 Export Date: 16 February 2020 Correspondence Address: Sankian, M.; Immuno-Biochemistry Lab., Immunology Research Center, Bu-Ali Research Institute, Ferdowsi Square, 919773117, Mashhad, Iran
Uncontrolled Keywords: Eletroporation Lactobacilli Plasmid size Probiotic Bioengineering research Cell walls Electric field strength Electroporation Electrotransformation Food industries Genetic manipulations Genetic modifications New protocol Probiotic lactobacilli Probiotic products Probiotics Pulse durations Standard method Standard protocols Transformants Transformation efficiency Washing buffers Amino acids DNA Efficiency Electric fields Washing Bacilli Bacteria (microorganisms) Lactobacillus Lactobacillus rhamnosus
Subjects: QW Microbiology and Immunology
Divisions: Mashhad University of Medical Sciences
Depositing User: mr lib5 lib5
Date Deposited: 10 May 2020 16:26
Last Modified: 10 May 2020 16:26
URI: http://eprints.mums.ac.ir/id/eprint/18883

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