Easily applicable modifications to electroporation conditions improve the transformation efficiency rates for rough morphotypes of fast-growing mycobacteria

Víctor Campo-Pérez, Maria del Mar Cendra, Esther Julián*, Eduard Torrents*

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Electroporation is the most widely used and efficient method to transform mycobacteria. Through this technique, fast- and slow-growing mycobacteria with smooth and rough morphotypes have been successfully transformed. However, transformation efficiencies differ widely between species and strains. In this study, the smooth and rough morphotypes of Mycobacteroides abscessus and Mycolicibacterium brumae were used to improve current electroporation procedures for fast-growing rough mycobacteria. The focus was on minimizing three well-known and challenging limitations: the mycobacterial restriction-modification systems, which degrade foreign DNA; clump formation of electrocompetent cells before electroporation; and electrical discharges during pulse delivery, which were reduced by using salt-free DNA solution. Herein, different strategies are presented that successfully address these three limitations and clearly improve the electroporation efficiencies over the current procedures. The results demonstrated that combining the developed strategies during electroporation is highly recommended for the transformation of fast-growing rough mycobacteria.

Original languageEnglish
Pages (from-to)10-18
Number of pages9
JournalNew Biotechnology
Volume63
DOIs
Publication statusPublished - 25 Jul 2021

Keywords

  • Clump
  • Desalted DNA
  • Electroporation
  • Mycobacteria
  • Mycobacteroides abscessus
  • Mycolicibacterium brumae

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