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

Li M, Cui Y, Gan Z, Shi C, Shi X - Mar Drugs (2015)

Bottom Line: 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 Article: PubMed Central - PubMed

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.

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.

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The expression of Cppsy gene induced by light (A) and MeJA (B) at different time points in C. protothecoides CS-41. Data (mean ± SEM) are combined from three independent experiments. # indicates that the gene expression levels were significantly different from that at 0 h (p < 0.01).
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marinedrugs-13-06620-f005: The expression of Cppsy gene induced by light (A) and MeJA (B) at different time points in C. protothecoides CS-41. Data (mean ± SEM) are combined from three independent experiments. # indicates that the gene expression levels were significantly different from that at 0 h (p < 0.01).

Mentions: To investigate the effects of light and MeJA on the promoter inducibility at different time points, we analyzed Cppsy mRNA expression levels after light and MeJA treatment. The results show that on treatment with light, Cppsy gene expression increased by up to 36 times, compared with the dark, which indicates that Cppsy gene expression is up-regulated in response to light (Figure 5A). When treated with MeJA, Cppsy gene expression peaked at 10 h after treatment (Figure 5B). Therefore, the gene expression of Cppsy is significantly induced by treatment with light and MeJA (p < 0.01). These results confirm that the Cppsy promoter is induced by light and MeJA, and can be used as a candidate promoter element for the genetic modification of carotenoid biosynthesis in Chlorella, other algae, or higher plants.


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

Li M, Cui Y, Gan Z, Shi C, Shi X - Mar Drugs (2015)

The expression of Cppsy gene induced by light (A) and MeJA (B) at different time points in C. protothecoides CS-41. Data (mean ± SEM) are combined from three independent experiments. # indicates that the gene expression levels were significantly different from that at 0 h (p < 0.01).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663545&req=5

marinedrugs-13-06620-f005: The expression of Cppsy gene induced by light (A) and MeJA (B) at different time points in C. protothecoides CS-41. Data (mean ± SEM) are combined from three independent experiments. # indicates that the gene expression levels were significantly different from that at 0 h (p < 0.01).
Mentions: To investigate the effects of light and MeJA on the promoter inducibility at different time points, we analyzed Cppsy mRNA expression levels after light and MeJA treatment. The results show that on treatment with light, Cppsy gene expression increased by up to 36 times, compared with the dark, which indicates that Cppsy gene expression is up-regulated in response to light (Figure 5A). When treated with MeJA, Cppsy gene expression peaked at 10 h after treatment (Figure 5B). Therefore, the gene expression of Cppsy is significantly induced by treatment with light and MeJA (p < 0.01). These results confirm that the Cppsy promoter is induced by light and MeJA, and can be used as a candidate promoter element for the genetic modification of carotenoid biosynthesis in Chlorella, other algae, or higher plants.

Bottom Line: 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 Article: PubMed Central - PubMed

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.

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.

Show MeSH
Related in: MedlinePlus