Regulatory

Part:BBa_K4263000

Designed by: Hanlin Li   Group: iGEM22_SCUT-China   (2022-09-23)
Revision as of 16:05, 30 September 2022 by Hanlin Li (Talk | contribs)

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K4263000

P0547

Introduction

0547 promoter is a novol promising methanol inducible promoter in the yeast Pichia pastoris[1].

Characterization

In order to test the function of P0547, we construct "P0547-EGFP-terminator" (Figure 1). If P0547 is functional, we can test the fluorescence intensity of EGFP in supernatant samples obtained from the culture of recombinant P.pastoris GS115 strain.

Figure 1 Gene circuit of P0547-EGFP-terminator

Our results matched the general expected trend (Figure 2). After fermentation experiment in BMMY medium containing 1% methanol. The fluorescence intensity of the samples of recombinant P.pastoris GS115 containing the EGFP gene gradually increased over time, which is in line with literature description[1]. At the same time, we measured the growth curve of the strains.

Figure 2 Fluorescence intensity and OD600 absorbance of samples obtained at different time points from the culture of corresponding recombinant P.pastoris GS115 containing EGFP gene.

Reference

[1] Xu N, Zhu J, Zhu Q, et al. Identification and characterization of novel promoters for recombinant protein production in yeast Pichia pastoris[J]. Yeast, 2018,35(5):379-385.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 416
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 416
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 416
  • 1000
    COMPATIBLE WITH RFC[1000]



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