All-in-one construct for genome engineering using Cre-lox technology

Ana M. Mariscal, Luis González-González, Enrique Querol, Jaume Piñol

Research output: Contribution to journalArticleResearchpeer-review

8 Citations (Scopus)

Abstract

© The Author 2016. Published by Oxford University Press on behalf of Kazusa DNA Research Institute. Mycoplasma genitalium is an appealing model of a minimal cell and synthetic biology study, and it was one of the first organisms whose genome was fully sequenced and chemically synthesized. Despite its usefulness as a model organism, many genetic tools well established for other microorganisms are not currently available in mycoplasmas. We have developed several vectors to adapt the Cre-lox technology for genome engineering in M. genitalium, providing an all-in-one construct that could be also useful to obtain unmarked genetic modifications in many other slow growing microorganisms. This construct contains a modified promoter sequence based in TetR system that exhibits an enhanced control on Cre recombinase expression, virtually abolishing the presence of this recombinase in the absence of inducer. This allows to introduce the Cre recombinase gene and the desired genetic modification in a single transformation step. In addition, this inducible promoter may be a very promising tool for a wide range of molecular applications.
Original languageEnglish
Pages (from-to)263-270
JournalDNA Research
Volume23
Issue number3
DOIs
Publication statusPublished - 15 Jun 2016

Keywords

  • Cre-lox technology
  • TetR-TetO inducible promoters
  • genome engineering
  • mycoplasmas

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