Composite

Part:BBa_K3711056

Designed by: Jiacheng Shi   Group: iGEM21_HUST-China   (2021-10-03)


Panb1-α factor-crtE-AOX1 Terminator


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 124
    Illegal XhoI site found at 649
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1325
  • 1000
    COMPATIBLE WITH RFC[1000]


Description

This is a composite part for extracellular expression of Geranylgeranyl pyrophosphate synthase. crtE is expressed and participates in the production from Farnessee pyrophosphate (FPP) to Geranylgeranyl pyrophosphate (GGPP). Panb1 is a constitutive promoter in yeast, which is expressed under anaerobic conditions, while under aerobic conditions, Panb1, as a repression target of ROX1, is inhibited. When Panb1 initiates the expression, the signal peptide, α-factor, is used to express crtE outside of the cell.

Usage and Biology

crtE is derived from Erwinia, encodes Geranylgeranyl pyrophosphate synthase and participates in the synthesis of carotenoids. The early steps of carotenoid biosynthesis pathway include the synthesis of Geranylgeranyl pyrophosphate (GGPP), the condensation of two molecules of GGPP into octahydrolycopene and desaturation of octahydrolycopene into plant fluorene, β-carotene, protolycopene and lycopene. The crtE encodes Geranylgeranyl pyrophosphate synthase which synthesizes GGPP. Laboratory studies have shown that E.coli transformed with E.herbicola carotenoid synthesis gene could resist higher level of LTV radiation and phototoxic environment, indicating that the synthesis of carotenoid may be very important for the survival of E.herbicola in nature.

Molecular cloning

Fig1. Plasmid construction and colony PCR results of Panb1-α factor-4CL-AOX1 Terminator, Panb1-α factor-crtI-AOX1 Terminator, Panb1-α factor-crtB-AOX1 Terminator and Panb1-α factor-crtE-AOX1 Terminator transformed E.coli
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Categories
Parameters
None