[22] Directed mutagenesis in photosystem II: Analysis of the CP 47 protein

Add time:08/04/2019    Source:sciencedirect.com

Publisher SummaryThis chapter describes the techniques used in laboratory to introduce directed mutations into the chlorophyll-protein CP 47 of the cyanobacterium Synechocystis 6803. This protein is an integral membrane component of the proximal chlorophyll a antenna of PS II, which also interacts with the oxygen-evolving site. The chapter investigates the roles of the conserved, charged residues located principally in the large extrinsic loop E of CP 47, which appears to be lumenally exposed. This domain is examined using a variety of biochemical techniques and there exists a strong body of evidence indicating that this portion of CP 47 interacts with components required for oxygen evolution. Site-directed mutations were introduced into this region of CP 47 using the method of Kunkel. Random mutations directed against the large extrinsic loop domain of CP 47 are introduced in the chapter using the mutator strain of E. coli XL-1 Red.

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