Difference between revisions of "Part:BBa K2971002"
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<partinfo>BBa_K2971002 short</partinfo>
 
<partinfo>BBa_K2971002 short</partinfo>
  
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This part is the gene crtE (dr1395) from the extremophile Deinococcus radiodurans and encodes
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This part is the gene <i>crtB</i> (<i>dr0862</i>) from the extremophile <i>Deinococcus radiodurans</i> and encodes
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geranylgeranyl diphosphate synthase (CrtE). GGPPS catalyze the condensation of farnesyl
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phytoene synthase (CrtB). Phytoene synthase catalyzes the first reaction in the biosynthetic pathway of
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pyrophosphate (FPP) and isopentenyl pyrophosphate (IPP) into geranylgeranyl diphosphate (GGPP).
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carotenoids [1]. The sequence has been codon optimized for expression in Escherichia coli and iGEM
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GGPP is a precursor for the carotenoid biosynthetic pathway. The sequence has been codon optimized
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registry compatibility.
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for expression in Escherichia coli and iGEM registry compatibility.
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The UiOslo 2019 team used this gene in a composite part to produce the red pigment lycopene in
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Escherichia coli by expressing it in a vector along with the genes crtE and crtI.
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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.
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'''References'''
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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
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Tian, B., & Hua, Y. (2010). Carotenoid biosynthesis in extremophilic Deinococcus&#x2013;Thermus bacteria. Trends in Microbiology, 18(11), 512-520. doi:10.1016/j.tim.2010.07.007
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 +
 
  
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The UiOslo 2019 team used this gene in a composite part (BBa_K2971004) to produce the red pigment lycopene, in
 
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Escherichia coli, by expressing it in a vector along with the genes crtB and crtI. The enzyme was put under the arabinose inducible promotor araC.
 
  
  

Revision as of 20:37, 21 October 2019


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 609
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]