Genetic modification of Cyanobacteria by conjugation using the CyanoGate modular cloning toolkit

Grant A. R. Gale, Alejandra A. Schiavon Osorio, Anton Puzorjov, Baojun Wang, Alistair J. Mccormick

Research output: Contribution to journalArticlepeer-review

Abstract

Cyanobacteria are a diverse group of prokaryotic photosynthetic organisms that can be genetically modified for the renewable production of useful industrial commodities. Recent advances in synthetic biology have led to development of several cloning toolkits such as CyanoGate, a standardized modular cloning system for building plasmid vectors for subsequent transformation or conjugal transfer into cyanobacteria. Here we outline a detailed method for assembling a self-replicating vector (e.g., carrying a fluorescent marker expression cassette) and conjugal transfer of the vector into the cyanobacterial strains Synechocystis sp. PCC 6803 or Synechococcus elongatus UTEX 2973. In addition, we outline how to characterize the performance of a genetic part (e.g., a promoter) using a plate reader or flow cytometry.
Original languageEnglish
Article numbere60451
Number of pages17
JournalJournal of Visualized Experiments
Issue number152
DOIs
Publication statusPublished - 31 Oct 2019

Keywords

  • flow cytometry
  • fluorescence
  • Golden Gate
  • plate reader
  • Synechocystis sp. PCC 6803
  • Synechococcus elongatus UTEX 2973
  • toolkit
  • transient expression

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