Efficient Heterologous Transformation of Chlamydomonas reinhardtii npq2 Mutant with the Zeaxanthin Epoxidase Gene Isolated and Characterized from Chlorella zofingiensisReport as inadecuate




Efficient Heterologous Transformation of Chlamydomonas reinhardtii npq2 Mutant with the Zeaxanthin Epoxidase Gene Isolated and Characterized from Chlorella zofingiensis - Download this document for free, or read online. Document in PDF available to download.

Institute of Plant Biochemistry and Photosynthesis, CIC Cartuja, University of Seville and CSIC, Avda. Américo Vespucio no. 49, 41092-Seville, Spain





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Abstract 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 Chlorella zofingiensis Czzep has been isolated. This gene encodes a polypeptide of 596 amino acids. A single copy of Czzep has been found in the C. zofingiensis genome by Southern blot analysis. qPCR analysis has shown that transcript levels of Czzep were increased after zeaxanthin formation under high light conditions. The functionality of Czzep gene by heterologous genetic complementation in the Chlamydomonas mutant npq2, which lacks zeaxanthin epoxidase ZEP activity and accumulates zeaxanthin in all conditions, was analyzed. The Czzep gene was adequately inserted in the pSI105 vector and expressed in npq2. 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 Chlamydomonas can be an efficient tool for heterologous expression and metabolic engineering for biotechnological applications. View Full-Text

Keywords: Chlorella zofingiensis; Chlamydomonas reinhardtii; zeaxanthin epoxidase; violaxanthin cycle; carotenoids Chlorella zofingiensis; Chlamydomonas reinhardtii; zeaxanthin epoxidase; violaxanthin cycle; carotenoids





Author: Inmaculada Couso, Baldo F. Cordero, María Ángeles Vargas and Herminia Rodríguez *

Source: http://mdpi.com/



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