The psbK gene encodes a small protein of Photosystem II. The gene has previously been cloned and sequenced in Synechocystis sp. PCC 6803. Our new results, presented here, confirm the conclusions of Ikeuchi et al. Based on Northern hybridization and primer extension analyses, we show that psbK is transcribed as a monocistronic message in this cyanobacterium. Analysis of DNA sequence immediately upstream of the transcription start site revealed an E. coli-like-10 consensus sequence. A deletion mutant was constructed where the psbK gene was replaced by a kanamycin resistant cartridge. In situ complementation experiments, as well as Southern and Northern hybridization analyses, confirmed that the mutant strain contains a lesion in psbK. The psbK-less mutant could grow photoautotrophically as well as photoheterotrophically both in liquid culture and on agar plates. The rate of growth was slightly less compared with the wild-type as clearly observed by in situ complementation experiments. Although the mutant showed correspondingly lower rates of electron transport, thermoluminescence, oxygen flash yield and chlorophyll a fluorescence measurements did not detect any significant modification of the reactions of PS II. Moreover, the mutant was no more susceptible to excess light than the wild-type. It is, therefore, concluded that the product of the psbK gene is not crucial for PS II activity and possibly plays some other role in the metabolism of Synechocystis.