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Molecular and General Genetics MGG

, Volume 216, Issue 1, pp 51–59 | Cite as

Mutagenesis by random cloning of an Escherichia coli kanamycin resistance gene into the genome of the cyanobacterium Synechocystis PCC 6803: Selection of mutants defective in photosynthesis

  • F. Chauvat
  • P. Rouet
  • H. Bottin
  • A. Boussac
Article

Summary

Photosynthetic mutants of the cyanobacterium Synechocystis PCC 6803 were produced by a random cartridge mutagenesis method leading to gene inactivation. This procedure relies on random ligation of an Escherichia coli kanamycin resistance (Kmr) gene to restriction fragments of genomic DNA from the host. Then recombination occurring during transformation promotes integration of the marker gene into the genome of the recipient cells. Several mutants impaired in photosynthesis were obtained by this procedure. All are partially or totally defective in photosystem II activity and some of them also harbour a functionally modified photosystem I. Restriction and recombination data showed that one mutant (AK1) is best explained as an insertion of the Kmr gene into an AvaII restriction site of the gene psbD-1. All other harbour a deletion, ranging from at least 1.15 kb (AK3) to more than 50 kb (AK9), which partly or fully overlaps the genes psbB and/or psbD-1, depending on the mutant. A genetic-physical map of the more than 60 kb region of the cyanobacterial genome harbouring the genes psbB, psbC and psbD-1 was constructed by combining published sequence data on these genes with the results of recombination and restriction mapping.

Key words

Random cartridge mutagenesis Transformation Recombination Genetic mapping 

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • F. Chauvat
    • 1
  • P. Rouet
    • 1
  • H. Bottin
    • 2
  • A. Boussac
    • 2
  1. 1.Service de BiochimieDepartement de Biologie, F-CEN SaclayGif-sur Yvette cedexFrance
  2. 2.Service de BiophysiqueDepartement de Biologie, F-CEN SaclayGif-sur Yvette cedexFrance

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