title: Use of Nicotiana tabacum transplastomic plants engineered to express a His-tagged CP47 for the isolation of functional photosystem II core complexes.
abstract: This work focuses on the development of a molecular tool for purification of Photosystem II (PSII) 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.
authors.affiliation: Applied Science and Technology Department-BioSolar Lab, Politecnico di Torino, Viale Teresa Michel 5, 15121 Alessandria, Italy. Electronic address: cristina.pagliano@polito.it.
authors.forename: Cristina
authors.initials: C
authors.lastname: Pagliano
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bersanini
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.
authors.forename: Rino
authors.initials: R
authors.lastname: Cella
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Longoni
authors.affiliation: Department of Biology and Biotechnology, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Pantaleoni
authors.affiliation: Plant Transformation Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Dass
authors.affiliation: Plant Transformation Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Sadhu
authors.initials: S
authors.lastname: Leelavathi
authors.affiliation: Plant Transformation Group, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India. Electronic address: vsreddy@icgeb.res.in.
authors.forename: Vanga Siva
authors.initials: VS
authors.lastname: Reddy
mesh_terms: D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D017343:Genes, Plant; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes / Q000302:isolation & purification* / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D045332:Photosystem II Protein Complex / Q000302:isolation & purification* / Q000378:metabolism; D030821:Plants, Genetically Modified; D018087:Plastids / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013057:Spectrum Analysis; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
chemical_list: D045342:Light-Harvesting Protein Complexes; D045332:Photosystem II Protein Complex; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; C070970:photosystem II, chlorophyll-binding protein, CP-47
keywords: Biolistic chloroplast transformation; CP47 His-tag; Photosystem II; Transplastomic tobacco plants