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Purification of His6-tagged Photosystem I from Chlamydomonas reinhardtii.
- Source :
- Photosynthesis Research; Apr2008, Vol. 96 Issue 1, p51-60, 10p
- Publication Year :
- 2008
-
Abstract
- Abstract  We have developed a rapid method for isolation of the Photosystem I (PS1) complex from Chlamydomonas reinhardtii using epitope tagging. Six histidine residues were genetically added to the N-terminus of the PsaA core subunit of PS1. The His6-tagged PS1 could be purified with a yield of 80â90% from detergent-solubilized thylakoid membranes within 3 h in a single step using a Ni-nitrilotriacetic acid (Ni-NTA) column. Immunoblots and low-temperature fluorescence analysis indicated that the His6-tagged PS1 preparation was highly pure and extremely low in uncoupled pigments. Moreover, the introduced tag appeared to have no adverse effect upon PS1 structure/function, as judged by photochemical assays and EPR spectroscopy of isolated particles, as well as photosynthetic growth tests of the tagged strain. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 01668595
- Volume :
- 96
- Issue :
- 1
- Database :
- Complementary Index
- Journal :
- Photosynthesis Research
- Publication Type :
- Academic Journal
- Accession number :
- 30080444
- Full Text :
- https://doi.org/10.1007/s11120-007-9283-9