Jump to main content
Jump to site search

Issue 11, 2007
Previous Article Next Article

A highly active histidine-tagged Chlamydomonas reinhardtii Photosystem II preparation for structural and biophysical analysis

Author affiliations

Abstract

We describe a genetically engineered strain of Chlamydomonas reinhardtii where the PsbH subunit of Photosystem II (PSII) has been modified to include a C-terminal polyhistidine tag. The strain was generated by the rescue to photoautotrophic growth of a psbH insertional mutant following chloroplast transformation with the modified gene. This selection strategy confirms that the addition of the tag to PsbH does not prevent the assembly of functional PSII, and results in an engineered strain with tagged PSII but no antibiotic-resistance markers in the chloroplast genome. Consequently, the strain is suitable for subsequent genetic manipulation of chloroplast PSII genes. We also describe a rapid PSII isolation procedure that gives a preparation capable of high rates of oxygen evolution. This preparation is suitable for spectroscopic analysis as shown by EPR analysis of the S2 state of the water oxidation cycle. Furthermore, electron microscopy, coupled to single particle analysis, has revealed the isolated PSII to be structurally homogeneous core dimers that are ideally suited for higher resolution structural studies.

Graphical abstract: A highly active histidine-tagged Chlamydomonas reinhardtii Photosystem II preparation for structural and biophysical analysis

Back to tab navigation

Article information


Submitted
06 Jun 2007
Accepted
21 Aug 2007
First published
03 Sep 2007

Photochem. Photobiol. Sci., 2007,6, 1177-1183
Article type
Paper

A highly active histidine-tagged Chlamydomonas reinhardtii Photosystem II preparation for structural and biophysical analysis

M. Cullen, N. Ray, S. Husain, J. Nugent, J. Nield and S. Purton, Photochem. Photobiol. Sci., 2007, 6, 1177
DOI: 10.1039/B708611N

Search articles by author

Spotlight

Advertisements