Stable expression and characterization of a fungal pectinase and bacterial peroxidase genes in tobacco chloroplast
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Espinoza Sánchez, Edward Alexander; Álvarez Hernández, Marianela Hazel; Torres Castillo, Jorge Ariel; Rascón Cruz, Quintín; Gutiérrez Díez, Adriana; [et al.]Patrocinadores
This work was financially supported by the Consejo Nacional de Ciencia y Tecnología (CONACYT) through the project CB-2012-01 179794. We would like to thank Dr. Pal Maliga, Waksman Institute, Rutgers University, USA for providing pPRV111A plasmid and Massimo Bosacchi for his critical reading of the manuscript and helpful comments.Fecha de publicación
2015Editorial
Pontificia Universidad Católica de ValparaísoCita bibliográfica
ESPINOZA-SANCHEZ, Edward Alexander et al. Stable expression and characterization of a fungal pectinase and bacterial peroxidase genes in tobacco chloroplast. Electron. J. Biotechnol. [online]. 2015, vol.18, n.3, pp.161-168. ISSN 0717-3458. http://dx.doi.org/10.1016/j.ejbt.2015.03.002.Revisión por pares
SiPalabras clave
Cell wallChloroplast genetic transformation
Hydrolytic enzyme
Nicotiana tabacum
Resumen
Background: The high capacity of chloroplast genome response to integrate and express transgenes at high levels makes this technology a good option to produce proteins of interest. This report presents the stable expression of Pectin lyase (PelA gene) and the first stable expression of manganese peroxidase (MnP-2 gene) from the chloroplast genome. Results: pES4 and pES5 vectors were derived from pPV111A plasmid and contain the PelA and MnP-2 synthetic genes, respectively. Both genes are flanked by a synthetic rrn16S promoter and the 3′UTR from rbcL gene. Efficient gene integration into both inverted repeats of the intergenic region between rrn16S and 3′rps′12 was confirmed by Southern blot. Stable processing and expression of the RNA were confirmed by Northern blot analysis. Enzymatic activity was evaluated to detect expression and functionality of both enzymes. In general, mature plants showed more activity than young transplastomic plants. Compared to wild type plants, transplastomic ...
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