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Circadian Rhythms

Volume 362 of the series Methods in Molecular Biology™ pp 155-171

Specialized Techniques for Site-Directed Mutagenesis in Cyanobacteria

  • Eugenia M. ClericoAffiliated withDepartment of Biology, Texas A&M University
  • , Jayna L. DittyAffiliated withDepartment of Biology
  • , Susan S. GoldenAffiliated withDepartment of Biology, Texas A&M University

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Abstract

Synechococcus elongatus PCC 7942 is an excellent model system for studying the molecular mechanism of the circadian clock in cyanobacteria. The “plastic” genetic characteristics of this organism have facilitated the development of various methods for mutagenesis of its chromosome. These methods are based on homologous recombination between the chromosome and foreign DNA, introduced to the cyanobacteria by either transformation or conjugation. Here we describe different approaches to mutagenize the chromosome of S. elongatus, including insertional mutagenesis, hit-and-run allele replacement, rps12-mediated gene replacement, and regulated expression of genes from ectopic sites, the neutral sites of the S. elongatus genome.

Key Words

Synechococcus elongatus PCC 7942 hit-and-run allele replacement in-frame deletion rps12-mediated gene replacement homologous recombination transformation neutral site