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Mar. Drugs 2015, 13(11), 6620-6635; doi:10.3390/md13116620

Isolation and Analysis of the Cppsy Gene and Promoter from Chlorella protothecoides CS-41

1,2,†
,
1,†
,
1
,
1,* and 1
1
MOST-USDA Joint Research Center for Food Safety, School of Agriculture and Biology, and State Key Lab of Microbial Metabolism, Shanghai Jiao Tong University, Shanghai 200240, China
2
Analytical Testing Center, Zhejiang Chinese Medical University, Hangzhou 310053, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Graziano Riccioni
Received: 25 June 2015 / Revised: 9 September 2015 / Accepted: 9 September 2015 / Published: 28 October 2015
(This article belongs to the Special Issue Marine Carotenoids (Special Issue))
View Full-Text   |   Download PDF [1117 KB, uploaded 28 October 2015]   |  

Abstract

Phytoene synthase (PSY) catalyzes the condensation of two molecules of geranylgeranyl pyrophosphate to form phytoene, the first colorless carotene in the carotenoid biosynthesis pathway. So it is regarded as the crucial enzyme for carotenoid production, and has unsurprisingly been involved in genetic engineering studies of carotenoid production. In this study, the psy gene from Chlorella protothecoides CS-41, designated Cppsy, was cloned using rapid amplification of cDNA ends. The full-length DNA was 2488 bp, and the corresponding cDNA was 1143 bp, which encoded 380 amino acids. Computational analysis suggested that this protein belongs to the Isoprenoid_Biosyn_C1 superfamily. It contained the consensus sequence, including three predicted substrate-Mg2+ binding sites. The Cppsy gene promoter was also cloned and characterized. Analysis revealed several candidate motifs for the promoter, which exhibited light- and methyl jasmonate (MeJA)-responsive characteristics, as well as some typical domains universally discovered in promoter sequences, such as the TATA-box and CAAT-box. Light- and MeJA treatment showed that the Cppsy expression level was significantly enhanced by light and MeJA. These results provide a basis for genetically modifying the carotenoid biosynthesis pathway in C. protothecoides. View Full-Text
Keywords: Chlorella protothecoides CS-41; phytoene synthase; Cppsy; promoter Chlorella protothecoides CS-41; phytoene synthase; Cppsy; promoter
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Li, M.; Cui, Y.; Gan, Z.; Shi, C.; Shi, X. Isolation and Analysis of the Cppsy Gene and Promoter from Chlorella protothecoides CS-41. Mar. Drugs 2015, 13, 6620-6635.

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