title: Isolation and Analysis of the Cppsy Gene and Promoter from Chlorella protothecoides CS-41.
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-Mg(2+) 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.
authors.affiliation: 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. lmeiya@126.com.|Analytical Testing Center, Zhejiang Chinese Medical University, Hangzhou 310053, China. lmeiya@126.com.
authors.forename: Meiya
authors.initials: M
authors.lastname: Li
authors.affiliation: 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. cyan9028@163.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Cui
authors.affiliation: 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. forrestgzb@yahoo.cn.
authors.forename: Zhibing
authors.initials: Z
authors.lastname: Gan
authors.affiliation: 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. clshi@sjtu.edu.cn.
authors.forename: Chunlei
authors.initials: C
authors.lastname: Shi
authors.affiliation: 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. xmshi@sjtu.edu.cn.
authors.forename: Xianming
authors.initials: X
authors.lastname: Shi
mesh_terms: D000085:Acetates / Q000494:pharmacology; D002708:Chlorella / Q000235:genetics*; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics; D005786:Gene Expression Regulation / Q000235:genetics; D005818:Genetic Engineering; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase / Q000235:genetics* / Q000302:isolation & purification; D008027:Light; D054883:Oxylipins / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics*
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D018076:DNA, Complementary; D054883:Oxylipins; C072239:methyl jasmonate; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase
keywords: Chlorella protothecoides CS-41; Cppsy; phytoene synthase; promoter