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Plant Physiology 85:1021-1025 (1987)
© 1987 American Society of Plant Biologists

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Molecular Biology and Gene Regulation

Construction of an Obligate Photoheterotrophic Mutant of the Cyanobacterium Synechocystis 6803 1

Inactivation of the psbA Gene Family

Christer Jansson, Richard J. Debus, Heinz D. Osiewacz, Michael Gurevitz and Lee McIntosh

MSU-DOE Plant Research Laboratory and Department of Biochemistry, Michigan State University, East Lansing, Michigan 48824, Lehrstuhl fur Allgemeine Botanik, Ruhr-Universität, Bochum, West Germany, Department of Botany, University of Tel Aviv, Tel Aviv, Israel

psbA in Synechocystis 6803 was found to belong to a small multigene family with three copies. The psbA gene family was inactivated in vitro by insertation of bacterial drug resistance markers. Inactivation of all three genes resulted in a transformant that is unable to grow photosynthetically but can be cultured photoheterotrophically. This mutant lacks oxygen evolving capacity but retains photosystem I activity. Room temperature measurements of chlorophyll a fluorescence induction demonstrated that the transformant exhibits a high fluorescence yield with little or no variable fluorescence. Immunoblot analyses showed complete loss of the psbA gene product (the DI polypeptide) from thylakoid membranes in the transformant. However, the extrinsic 33 kilodalton polypeptide of the water-splitting complex of photosystem II, is still present. The results indicate that assembly of a partial photosystem II complex may occur even in the absence of the intrinsic D1 polypeptide, a protein implicated as a crucial component of the photosystem II reaction center.

1 Supported by the McKnight Foundation (C. J., R. J. D.); The Swedish National Science Research Council (C. J.); National Science Foundation Postdoctoral Fellowship Grant DMB-8608566 (R. J. D.); The Deutsche Forschungsgemeinschaft (H. D. O.); and Department of Energy Contract DE-AC02-76ERO-1338.




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