Purification of His6-tagged Photosystem I from Chlamydomonas reinhardtii View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2008-01-04

AUTHORS

Galina Gulis, Kuppala V. Narasimhulu, Lisa N. Fox, Kevin E. Redding

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 80–90% 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. More... »

PAGES

51-60

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s11120-007-9283-9

DOI

http://dx.doi.org/10.1007/s11120-007-9283-9

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1026230771

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/18175204


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