Genetic engineering of the green alga Chlorella zofingiensis: a modified norflurazon-resistant phytoene desaturase gene as a dominant selectable marker
Liu, Jin1,2,7; Sun, Zheng3; Gerken, Henri4; Huang, Junchao5; Jiang, Yue6; Chen, Feng1,7; Liu,J (reprint author),Peking Univ,Coll Engn,Inst Food & Bioresource Engn,Beijing 100871,Peoples R China.; jliu@umces.edu; sfchencoe@pku.edu.cn
2014-06-01
发表期刊APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
ISSN0175-7598
卷号98期号:11页码:5069-5079
摘要The unicellular green alga Chlorella zofingiensis has been proposed as a promising producer of natural astaxanthin, a commercially important ketocarotenoid. But the genetic toolbox for this alga is not available. In the present study, an efficient transformation system was established for C. zofingiensis. The transformation system utilized a modified norflurazon-resistant phytoene desaturase (PDS-L516F, with an leucine-phenylalanine change at position 516) as the selectable marker. Three promoters from endogenous PDS, nitrate reductase (NIT), and ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RBCS) genes were tested, with the RBCS promoter demonstrating the highest transformation efficiency. Inclusion of the first intron of the PDS gene further enhanced the efficiency by 91 %. Both particle bombardment and electroporation methods were examined, and the latter gave a fourfold higher transformation efficiency. The introduction of PDS-L516F, which exhibited a 33 % higher desaturation activity than the unaltered enzyme, enabled C. zofingiensis to produce 32.1 % more total carotenoids (TCs) and 54.1 % more astaxanthin. The enhanced accumulation of astaxanthin in transformants was revealed to be related to the increase in the transcripts of PDS, beta-carotenoid ketolase (BKT), and hydroxylase (CHYb) genes. Our study clearly shows that the modified PDS gene is a dominant selectable marker for the transformation of C. zofingiensis and possibly for the genetic engineering of the carotenoid biosynthetic pathway. In addition, the engineered C. zofingiensis might serve as an improved source of natural astaxanthin.
关键词Astaxanthin Chlorella Zofingiensis Genetic Engineering Norflurazon Phytoene Desaturase Transformation
资助信息985 Project of Peking University;State Oceanic Administration of China;National Research Foundation of Singapore
收录类别SCI
语种英语
引用统计
文献类型期刊论文
条目标识符http://ir.kib.ac.cn/handle/151853/18214
专题资源植物与生物技术所级重点实验室
通讯作者Liu,J (reprint author),Peking Univ,Coll Engn,Inst Food & Bioresource Engn,Beijing 100871,Peoples R China.; jliu@umces.edu; sfchencoe@pku.edu.cn
作者单位1.Peking Univ, Coll Engn, Inst Food & Bioresource Engn, Beijing 100871, Peoples R China
2.Univ Maryland, Ctr Environm Sci, Inst Marine & Environm Technol, Baltimore, MD 21201 USA
3.Shanghai Ocean Univ, Coll Fisheries & Life Sci, Shanghai, Peoples R China
4.Arizona State Univ, Dept Appl Sci & Math, Mesa, AZ USA
5.Chinese Acad Sci, Kunming Inst Bot, Kunming, Peoples R China
6.Jiangnan Univ, Sch Food Sci, Wuxi, Peoples R China
7.Singapore Peking Univ Res Ctr Sustainable Low Car, Singapore, Singapore
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Liu, Jin,Sun, Zheng,Gerken, Henri,et al. Genetic engineering of the green alga Chlorella zofingiensis: a modified norflurazon-resistant phytoene desaturase gene as a dominant selectable marker[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2014,98(11):5069-5079.
APA Liu, Jin.,Sun, Zheng.,Gerken, Henri.,Huang, Junchao.,Jiang, Yue.,...&sfchencoe@pku.edu.cn.(2014).Genetic engineering of the green alga Chlorella zofingiensis: a modified norflurazon-resistant phytoene desaturase gene as a dominant selectable marker.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,98(11),5069-5079.
MLA Liu, Jin,et al."Genetic engineering of the green alga Chlorella zofingiensis: a modified norflurazon-resistant phytoene desaturase gene as a dominant selectable marker".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 98.11(2014):5069-5079.
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