A1 Refereed original research article in a scientific journal
Use of Nicotiana tabacum transplastomic plants engineered to express a His-tagged CP47 for the isolation of functional photosystem II core complexes
Authors: Pagliano C, Bersanini L, Cella R, Longoni P, Pantaleoni L, Dass A, Leelavathi S, Reddy VS
Publisher: ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER
Publication year: 2017
Journal: Plant Physiology and Biochemistry
Journal name in source: PLANT PHYSIOLOGY AND BIOCHEMISTRY
Journal acronym: PLANT PHYSIOL BIOCH
Volume: 111
First page : 266
Last page: 273
Number of pages: 8
ISSN: 0981-9428
eISSN: 1873-2690
DOI: https://doi.org/10.1016/j.plaphy.2016.12.009(external)
Abstract
This work focuses on the development of a molecular tool for purification of Photosystem II (MI) from Nicotiana tabacum (L). To this end, the chloroplast psbB gene encoding the CP47 PSII subunit was replaced with an engineered version of the same gene containing a C-terminal His-tag. Molecular analyses assessed the effective integration of the recombinant gene and its expression. Despite not exhibiting any obvious phenotype, the transplastomic plants remained heteroplasmic even after three rounds of regeneration under antibiotic selection. However, the recombinant His-tagged CP47 protein associated in vivo to the other PSII subunits allowing the isolation of a functional PSII core complex, although with low yield of extraction. These results will open up possible perspectives for further spectroscopic and structural studies. (C) 2016 Elsevier Masson SAS. All rights reserved.
This work focuses on the development of a molecular tool for purification of Photosystem II (MI) from Nicotiana tabacum (L). To this end, the chloroplast psbB gene encoding the CP47 PSII subunit was replaced with an engineered version of the same gene containing a C-terminal His-tag. Molecular analyses assessed the effective integration of the recombinant gene and its expression. Despite not exhibiting any obvious phenotype, the transplastomic plants remained heteroplasmic even after three rounds of regeneration under antibiotic selection. However, the recombinant His-tagged CP47 protein associated in vivo to the other PSII subunits allowing the isolation of a functional PSII core complex, although with low yield of extraction. These results will open up possible perspectives for further spectroscopic and structural studies. (C) 2016 Elsevier Masson SAS. All rights reserved.
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