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PLANT PHYSIOLOGY , Vol 111, Issue 4 1067-1075, Copyright © 1996 by American Society of Plant Biologists

GENE REGULATION AND MOLECULAR GENETICS

The Cyanobacterial Thioredoxin Gene Is Required for Both Photoautotrophic and Heterotrophic Growth

F. Navarro and F. J. Florencio
Instituto de Bioquimica Vegetal y Fotosintesis and Departamento de Bioquimica Vegetal y Biologia Molecular, Universidad de Sevilla-Consejo Superior de Investigaciones Cientificas, Apdo. 1113, 41080 Sevilla, Spain

The gene encoding thioredoxin in the facultative heterotrophic cyanobacterium Synechocystis sp. PCC 6803 (trxA) has been cloned by heterologous hybridization using the corresponding gene trxM from the cyanobacterium Anacystis nidulans as a probe. The deduced amino acid sequence of trxA predicts a protein of relative molecular weight of 11,750 and has strong identity with its cyanobacterial counterparts and other m-type thioredoxins of photosynthetic eukaryotes. The trxA gene has been expressed in Escherichia coli as a functional protein of 12 kD and has been shown by western blot analysis to be the same size as in Synechocystis. The trxA gene is transcribed in Synechocystis as a single transcript of 450 nucleotides and accumulates to the same level under photo synthetic and heterotrophic growth conditions. The trxA gene was inactivated with a kanamycin-resistant cassette, and total segregation of the disrupted trxA gene was obtained only when the trxM gene from A. nidulans (E.D.G. Muller, B.B. Buchanan [1989] J Biol Chem 264: 4008-4014) was simultaneously expressed in Synechocystis. Our results suggest an essential role of thioredoxin not only when cells grow photosynthetically but also under heterotrphic conditions.


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