crtI coding sequence encoding phytoene dehydrogenase
This is the coding sequence of crtI from Pantoea ananatis (formerly Erwinia uredovora) (Accession number D90087). It encodes phytoene dehydrogenase, part of the carotenoid biosynthesis pathway, which converts Phytoene to Lycopene (Misawa, N., Nakagawa, N., Kobayashi, K., Yamano, S., Nakamura, K., and Harashima, K. 1990. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. Journal of Bacteriology 172, 6704-612). Two PstI sites were removed by site-directed mutagenesis.
Improvement on BBa_K118003
BBa_K118003 is the coding sequence of phytoene desaturase (CrtI/PDS) in Erwinia uredovora. CrtI is an enzyme catalyzes the conversion of phytoene to lycopene in a one-step reaction. In iGEM16_SCAU-China project, CrtI is used to simplified a reconstructed pathway to achieve astaxanthin biosynthesis in higher plant, rice (Oryza sativa). The original CrtI is not suitable to express in higher plants. Thus, codon optimization on CrtI had made for rice. In addition, a Pea transit peptide of Rubisco small subunit is fused to the optimized CDS of CrtI for proper sorting into the plastid. Comparison of nucleaotide and amino acid sequence of original CrtI (EuCrtI) and codon-optimized CrtI (coCrtI) are shown in Figure.1 and Figure.2.
With the help of improved part, iGEM16_SCAU-China had successfully achieved astaxanthin biosynthesis in rice (Figure. 3). Content and purity of astaxanthin was determined by HPLC analysis (Figure. 4)
For more details about the design of optimized CrtI, please refer to http://parts.igem.org/Part:BBa_K1959002. For more details and analyses, please visit the wiki of SCAU-China 2016 (http://2016.igem.org/Team:SCAU-China).
[Edited by Junyu Chen, SCAU-China 2016, 19 October 2016(UTC)]
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21Illegal BamHI site found at 71
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC