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
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.