Team:Nankai/Measurement
Is measurement important?
Precise measurement is crucial since it is the foundation of every scientific discipline. More importantly, measurement can also be an act of communication if various laboratories around the world collaborate together and make meaningful comparisons between same or similar observations.
How can we help?
Strains capable for producing γ-PGA are divided into two categories based on their requirement for glutamate acid: glutamate-dependent strains and glutamate-independent strains. Glutamate-independent strains are preferable for industrial production because of their low cost and simplified fermentation process. However, compared with glutamate-dependent strains, their lower γ-PGA productivity limits their industrial application. Therefore, the construction of a glutamate-independent strain with high γ-PGA yield is important for industrial applications.
What did we do?
In our experiment, we use the coding gene of Green Fluorescence Protein as a new reporter gene. First we successfully constructed three recombinant plasmids, in which the three promoters are located upstream of the GFP coding gene. Then these recombinant plasmids were transformed into the E. coli cells and several strains that steadily express GFP were obtained. After the transformed strains were obtained, they were firstly observed under a fluorescent microscopy.Next, to confirm our result quantitatively, we used a flow cytometry to measure the average fluorescence intensity of each strain, as instructed.We also made a figure of the average fluorescence intensity to show the different transcription initiation ability of each promoter more clearly.
And the result?
The results are as following:
Fig.1 Fluorescent Microscopy Picture of Transformed E. coli
