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Construction of a psb27 mutagenesis system to study the role of Psb27 on oxygenic photosynthesis and the characterization of Synechocystis sp. PCC 6803 with a histidine tagged CP47, lacking photosystem I and phycobilisome function

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Construction of a psb27 mutagenesis system to study the role of Psb27 on oxygenic photosynthesis and the characterization of Synechocystis sp. PCC 6803 with a histidine tagged CP47, lacking photosystem I and phycobilisome function

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Samenvatting

Abstract

Oxygenic photosynthesis is the process where water splits into oxygen, protons and electrons. There are four multisubunit protein complexes that are involved in the process: photosystem II (PSII), cytochrome b6f, photosystem I (PSI) and ATPase. The complex that is responsible for the production of oxygen is PSII. During the reaction the D1 subunit of the complex gets damaged by photoinhibition. Psb27 is a lipoprotein and it has been hypothesized that this protein has a role in the repair cycle of PSII.

The aims of this study were: Construction of a new "clean" deletion plasmid and a mutagenesis system to study the effect of Psb27 on oxygenic photosynthesis,
characterization of Synechocystis sp. PCC 6803 PSI-/apcE-/CP47-His and study the effect of change in pH level.

The Δpsb27 deletion plasmid contained for the second time a deletion of approximately 200 bp at the same location. The same problem is noticed during the construction of the Δpsb27 deletion plasmid for the mutagenesis system. In this case there was no DNA deleted but the upstream and resistance cassette were ligated into the downstream plasmid seven times in the incorrect orientation. Two insertions where discovered after sequencing the mutagenesis plasmid, that need to be fixed.

The characterization experiments growth curve, whole cell absorption spectra, 77K fluorescence and oxygen evolution showed that the mutant strains are behaving as expected. pH experiments showed that the strains PSI-/apcE-/H469H and PSI-/apcE-/CP47-His can survive in pH 10, but are highly stressed at this condition.

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OrganisatieAvans Hogeschool
AfdelingATGM Academie voor de technologie van Gezondheid en Milieu
PartnersUniversity of Otago
TypeBachelor
TaalEngels

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