Engineering of an endogenous phytoene desaturase gene as a dominant selectable marker for Chlamydomonas reinhardtii transformation and enhanced biosynthesis of carotenoids

doi:10.1016/j.procbio.2013.04.020

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	Engineering of an endogenous phytoene desaturase gene as a dominant selectable marker for Chlamydomonas reinhardtii transformation and enhanced biosynthesis of carotenoids
	Liu, Jin 1,4; Gerken, Henri 2; Huang, Junchao3 ; Chen, Feng 4
刊名	PROCESS BIOCHEMISTRY
	2013-05-01
卷号	48 期号:5-6 页码:788-795
关键词	Chlamydomonas Reinhardtii Carotenoids Norflurazon Phytoene Desaturase Selectable Marker Site-directed Mutation
ISSN号	1359-5113
DOI	10.1016/j.procbio.2013.04.020
通讯作者	Liu, J (reprint author), Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA. ; jliu@umces.edu ; sfchencoe@pku.edu.cn
文献子类	Article
英文摘要	A phytoene desaturase (PDS) gene was cloned and characterized from the unicellular green microalga Chlamydomonas reinhardtii. Functional complementation analysis revealed C reinhardtii PDS (CrPDS) catalyzes the conversion of phytoene to the colored carotenoid zeta-carotene. A single amino acid substitution, L505F, enhanced its desaturation activity by 29%, as indicated by an in vitro enzymatic assay. In addition, CrPDS-L505F exhibited 27.7-fold higher resistance to the herbicide norflurazon. Glass bead-mediated delivery displayed a high transformation efficiency of C reinhardtii with CrPDS-L505F, demonstrating clearly that the engineered endogenous CrPDS is a dominant selectable marker for C. reinhardtii and possibly for other green algae. Furthermore, the expression of PDS could enhance the intracellular carotenoid accumulation of transformants, opening up the possibility of engineering the carotenogenic pathway for improved carotenoid production in microalgae. (C) 2013 Elsevier Ltd. All rights reserved.
学科主题	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering, Chemical
WOS关键词	NUCLEAR TRANSFORMATION ; CHLORELLA-ZOFINGIENSIS ; HERBICIDE NORFLURAZON ; FOREIGN GENE ; EXPRESSION ; RESISTANCE ; MUTANTS ; PLANTS ; PCR
WOS研究方向	Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology ; Engineering
语种	英语
WOS记录号	WOS:000321423600006
资助机构	985 Project of Peking University; State Oceanic Administration of China; 863 Project of the Ministry of Science and Technology of China
公开日期	2013-10-16
内容类型	期刊论文
源URL	[http://ir.kib.ac.cn:8080/handle/151853/16843]
专题	昆明植物研究所_资源植物与生物技术所级重点实验室
作者单位	1.Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA 2.Arizona State Univ, Dept Appl Sci & Math, Mesa, AZ USA 3.Chinese Acad Sci, Kunming Inst Bot, Kunming, Peoples R China 4.Peking Univ, Coll Engn, Inst Food & Bioresource Engn, Beijing 100871, Peoples R China
推荐引用方式 GB/T 7714	Liu, Jin,Gerken, Henri,Huang, Junchao,et al. Engineering of an endogenous phytoene desaturase gene as a dominant selectable marker for Chlamydomonas reinhardtii transformation and enhanced biosynthesis of carotenoids[J]. PROCESS BIOCHEMISTRY,2013,48(5-6):788-795.
APA	Liu, Jin,Gerken, Henri,Huang, Junchao,&Chen, Feng.(2013).Engineering of an endogenous phytoene desaturase gene as a dominant selectable marker for Chlamydomonas reinhardtii transformation and enhanced biosynthesis of carotenoids.PROCESS BIOCHEMISTRY,48(5-6),788-795.
MLA	Liu, Jin,et al."Engineering of an endogenous phytoene desaturase gene as a dominant selectable marker for Chlamydomonas reinhardtii transformation and enhanced biosynthesis of carotenoids".PROCESS BIOCHEMISTRY 48.5-6(2013):788-795.