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Título: | Efficient Heterologous Transformation of Chlamydomonas reinhardtii npq2 Mutant with the Zeaxanthin Epoxidase Gene Isolated and Characterized from Chlorella zofingiensis |
Autor: | Couso, Inmaculada CSIC ORCID ; Cordero, Baldo F. CSIC; Vargas, M. Ángeles CSIC; Rodríguez, Herminia CSIC | Fecha de publicación: | 12-sep-2012 | Editor: | Multidisciplinary Digital Publishing Institute | Citación: | Marine Drugs 10 (9): 1955-1976 (2012) | Resumen: | In the violaxanthin cycle, the violaxanthin de-epoxidase and zeaxanthin epoxidase catalyze the inter-conversion between violaxanthin and zeaxanthin in both plants and green algae. The zeaxanthin epoxidase gene from the green microalga <em>Chlorella zofingiensis</em> (<em>Czzep</em>) has been isolated<em>. </em>This gene encodes a polypeptide of 596 amino acids. A single copy of <em>Czzep</em> has been found in the <em>C. zofingiensis</em> genome by Southern blot analysis. qPCR analysis has shown that transcript levels of <em>Czzep</em> were increased after zeaxanthin formation under high light conditions. The functionality of <em>Czzep</em> gene by heterologous genetic complementation in the <em>Chlamydomonas</em> mutant <em>npq2</em>, which lacks zeaxanthin epoxidase<strong> (</strong>ZEP) activity and accumulates zeaxanthin in all conditions, was analyzed. The <em>Czzep</em> gene was adequately inserted in the pSI105 vector and expressed in <em>npq2</em>. The positive transformants were able to efficiently convert zeaxanthin into violaxanthin, as well as to restore their maximum quantum efficiency of the PSII (Fv/Fm). These results show that <em>Chlamydomonas</em> can be an efficient tool for heterologous expression and metabolic engineering for biotechnological applications. | Versión del editor: | http://doi.org/10.3390/md10091955 | URI: | http://hdl.handle.net/10261/156770 | DOI: | 10.3390/md10091955 | Identificadores: | doi: 10.3390/md10091955 |
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