PMID: 2498291Jun 1, 1989Paper

Insertional mutagenesis by random cloning of antibiotic resistance genes into the genome of the cyanobacterium Synechocystis strain PCC 6803

Journal of Bacteriology
J LabarreP Thuriaux

Abstract

The facultative heterotrophic cyanobacterium Synechocystis sp. strain PCC 6803 was transformed by HaeII Cmr fragments ligated at random to HaeII DNA fragments of the host genome. A similar transformation was done with an AvaII Kmr marker ligated to AvaII host DNA fragments. Integration of the resistance markers into the host genome led to a high frequency of stable Kmr and Cmr transformants. Physical analysis of individual transformants indicated that this result was due to homologous recombination by conversionlike events leading to insertion of the Cmr (or Kmr) gene between two HaeII (or AvaII) sites of the host genome, with precise deletion of the host DNA between these sites. In contrast, integrative crossover of circular DNA molecules with homology to the host DNA is very rare in this cyanobacterium. Strain PCC 6803 was shown to have about 12 genomic copies per cell in standard growth conditions, which complicates the detection of recessive mutations induced by chemical or UV mutagenesis. Random disruption of the host DNA by insertional transformation provides a convenient alternative to transposon mutagenesis in cyanobacteria and may help to overcome the difficulties encountered in generating recessive mutants by classica...Continue Reading

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Citations

Sep 10, 2009·Molecular Microbiology·Martial MarboutyFranck Chauvat
Nov 19, 2011·FEMS Microbiology Letters·Marco GrieseJörg Soppa
Oct 18, 2014·Frontiers in Bioengineering and Biotechnology·Daniel Camsund, Peter Lindblad
Jan 10, 2019·Biotechnology for Biofuels·Kamonchanock EungrasameeSaowarath Jantaro
May 1, 2020·Frontiers in Plant Science·Matthew A PopePeter J Nixon
Jan 13, 2017·BioMed Research International·Xiaohui ZhangFeng Luo
Aug 29, 2019·Frontiers in Microbiology·Xavier KammerscheitCorinne Cassier-Chauvat
Jan 20, 1999·Proceedings of the National Academy of Sciences of the United States of America·S ReumannK Keegstra
May 10, 2011·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Anna ZorinaDmitry A Los
Jan 30, 2013·Biochemical Society Transactions·Jörg Soppa
Nov 28, 2015·Biotechnology for Biofuels·Petter HammarHaakan N Joensson
Feb 12, 2010·Comparative and Functional Genomics·Mark J LawsonLiqing Zhang
Oct 21, 2005·Proceedings of the National Academy of Sciences of the United States of America·Mitsuhiro ItayaKyoko Fujita
May 8, 2004·Molecular Microbiology·Khalil MazouniFranck Chauvat
Dec 2, 2014·Applied Microbiology and Biotechnology·Kangming ChenRuanbao Zhou

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