Chimaeric CP47 mutants of the cyanobacterium Synechocystis sp. PCC 6803 carrying spinach sequences

Construction and function

Willem Vermaas, Gaozhong Shen, Itzhak Ohad

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Chimaeric mutants of the cyanobacterium Synechocystis sp. PCC 6803 have been generated carrying part or all of the spinach psbB gene, encoding CP47 (one of the chlorophyll-binding core antenna proteins in Photosystem II). The mutant in which the entire psbB gene had been replaced by the homologous gene from spinach was an obligate photoheterotroph and lacked Photosystem II complexes in its thylakoid membranes. However, this strain could be transformed with plasmids carrying selected regions of Synechocystis psbB to give rise to photoautotrophs with a chimaeric spinach/cyanobacterial CP47 protein. This process involved heterologous recombination in the cyanobacterium between psbB sequences from spinach and Synechocystis 6803, which was found to be reasonably effective in Synechocystis. Also other approaches were used that can produce a broad spectrum of chimaeric mutants in a single experiment. Functional characterization of the chimaeric photoautotrophic mutants indicated that if a decrease in the photoautotrophic growth rates was observed, this was correlated with a decrease in the number of Photosystem II reaction centers (on a chlorophyll basis) in the thylakoid membrane and with a decrease in oxygen evolution rates. Remaining Photosystem II reaction centers in these chimaeric mutants appeared to function rather normally, but thermoluminescence and chlorophyll a fluorescence measurements provided evidence for a destabilization of Q(B)/-. This illustrates the sensitivity of the functional properties of the PS II reaction center to mild perturbations in a neighboring protein.

Original languageEnglish (US)
Pages (from-to)147-162
Number of pages16
JournalPhotosynthesis Research
Volume48
Issue number1-2
StatePublished - 1996

Fingerprint

Synechocystis sp. PCC 6803
Synechocystis
Photosystem II Protein Complex
Spinacia oleracea
Cyanobacteria
spinach
photosystem II
mutants
Thylakoids
Chlorophyll
chlorophyll
thylakoids
Genes
Membranes
photoheterotrophs
Proteins
Thermoluminescence
Gene encoding
genes
proteins

Keywords

  • gene replacement
  • photosynthesis
  • Photosystem II
  • thylakoid membranes
  • transformation

ASJC Scopus subject areas

  • Plant Science

Cite this

Chimaeric CP47 mutants of the cyanobacterium Synechocystis sp. PCC 6803 carrying spinach sequences : Construction and function. / Vermaas, Willem; Shen, Gaozhong; Ohad, Itzhak.

In: Photosynthesis Research, Vol. 48, No. 1-2, 1996, p. 147-162.

Research output: Contribution to journalArticle

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