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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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Related in: MedlinePlus

Promoter sequence of Cppsy from C. protothecoides CS-41. Numbers indicate the positions relative to transcriptional start site (TSS). The TSS is indicated as +1 and in bold; important cis-elements are underlined.
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marinedrugs-13-06620-f004: Promoter sequence of Cppsy from C. protothecoides CS-41. Numbers indicate the positions relative to transcriptional start site (TSS). The TSS is indicated as +1 and in bold; important cis-elements are underlined.

Mentions: The promoter region of the Cppsy gene was cloned from C. protothecoides CS-41 genomic DNA. The cloned Cppsy promoter region was determined to be 1980 bp in length, and the sequence is shown in Figure 4. Furthermore, the cloned Cppsy promoter region was analyzed using the PLACE and PlantCARE databases. Several core fragments were identified, which are homologous to the cis-acting elements of higher plants and of great importance for the promoter functions (Figure 4). Three types of elements, which have been found to be regulated by hormones in the upstream region of some plant genes, are present in the Cppsy promoter: the ABRE type (CCTGCGTGGC, CACGTG, and GCCTCGTGGC) involved in abscisic acid responsiveness; the TGACG-motif (TGACG), the cis-acting regulatory element involved in methyl jasmonate (MeJA) responsiveness; and the Sp1 (CCCCCGCCA and ACCCGCCATG), MNF (GTGCCCCATGCAGGTT) and Box I (TTTCAAA) types involved in light responsiveness.


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)

Promoter sequence of Cppsy from C. protothecoides CS-41. Numbers indicate the positions relative to transcriptional start site (TSS). The TSS is indicated as +1 and in bold; important cis-elements are underlined.
© Copyright Policy
Related In: Results  -  Collection

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

marinedrugs-13-06620-f004: Promoter sequence of Cppsy from C. protothecoides CS-41. Numbers indicate the positions relative to transcriptional start site (TSS). The TSS is indicated as +1 and in bold; important cis-elements are underlined.
Mentions: The promoter region of the Cppsy gene was cloned from C. protothecoides CS-41 genomic DNA. The cloned Cppsy promoter region was determined to be 1980 bp in length, and the sequence is shown in Figure 4. Furthermore, the cloned Cppsy promoter region was analyzed using the PLACE and PlantCARE databases. Several core fragments were identified, which are homologous to the cis-acting elements of higher plants and of great importance for the promoter functions (Figure 4). Three types of elements, which have been found to be regulated by hormones in the upstream region of some plant genes, are present in the Cppsy promoter: the ABRE type (CCTGCGTGGC, CACGTG, and GCCTCGTGGC) involved in abscisic acid responsiveness; the TGACG-motif (TGACG), the cis-acting regulatory element involved in methyl jasmonate (MeJA) responsiveness; and the Sp1 (CCCCCGCCA and ACCCGCCATG), MNF (GTGCCCCATGCAGGTT) and Box I (TTTCAAA) types involved in light responsiveness.

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