Construction of a psb C deletion strain in Synechocystis 6803.
Synechocystis 6803 is a cyanobacterium that carries out-oxygenic photosynthesis. We are interested in the introduction of mutations in the large extrinsic loop region of the CP43 protein of Photosystem II (PSII). CP43 appears to be required for the stable assembly of the PSII complex and also appears to play a role in photosynthetic oxygen evolution. Deletion of short segments of the large extrinsic loop results in mutants incapable of evolving oxygen. Alterations in psbC, the gene encoding CP43, are introduced into Synechocystis 6803 by transformation and homologous recombination. Specifically, plasmid constructs bearing the site-directed mutations are introduced into a deletion strain where the portion of the gene encoding the area of mutation has been deleted and replaced by a gene conferring antibiotic resistance. We have constructed a deletion strain of Synechocystis appropriate for the introduction of mutations in the large extrinsic loop of CP43 and have used it successfully to produce site-directed mutants.
Goldfarb
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N
Department of Biology, East Carolina University, Greenville, NC 27858, USA.
Knoepfle
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N
Putnam-Evans
C
C
D000458:Cyanobacteria / Q000235:genetics* / Q000378:metabolism; D004579:Electron Transport; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D016297:Mutagenesis, Site-Directed; D010100:Oxygen / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000235:genetics*; D045332:Photosystem II Protein Complex; D015183:Restriction Mapping
D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; C083100:photosystem II, chlorophyll binding protein, CP-43; D010100:Oxygen