Part:BBa_K2971002
crtB from Deinococcus radiodurans
This part is the gene crtB (dr0862) from the extremophile Deinococcus radiodurans and encodes phytoene synthase (CrtB). Phytoene synthase catalyzes the first reaction in the biosynthetic pathway of carotenoids [1]. The sequence has been codon optimized for expression in Escherichia coli and iGEM registry compatibility.
The UiOslo 2019 team used this gene in a composite part to produce the red pigment lycopene in Escherichia coli by expressing it in a vector along with the genes crtE and crtI.
We had the gene synthesized for free by IDT. The gene was cloned into an expression vector under the control of arabinose inducible promotor araC. The cloning was done via the gibson method. Successful insertion was confirmed with colony PCR of transformed colonies (figure 1), followed by sequencing to check for any potential mutation or frameshift.
References
1. Liu, C., Sun, Z., Shen, S., Lin, L., Li, T., Tian, B., & Hua, Y. (2014). Identification and characterization of the geranylgeranyl diphosphate synthase in Deinococcus radiodurans. Letters in Applied Microbiology, 58(3), 219-224. doi:10.1111/lam.12181 Tian, B., & Hua, Y. (2010). Carotenoid biosynthesis in extremophilic Deinococcus–Thermus bacteria. Trends in Microbiology, 18(11), 512-520. doi:10.1016/j.tim.2010.07.007
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 609
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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