PPGK1
Introduction
PGK1 promoter is the promoter for the glycolytic enzyme 3-phosphoglycerate kinase[1].
Characterization
In order to test the function of PPGK1, we construct "PPGK1-EGFP-terminator" (Figure 1). If PPGK1 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 PPGK1-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 decreased during 24-72h. 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] Stadlmayr G, Mecklenbrauker A, Rothmuller M, et al. Identification and characterisation of novel Pichia pastoris promoters for heterologous protein production[J]. J Biotechnol, 2010,150(4):519-529.