Results were divided into three categories, similar to the experiments. More specifically, these categories are Preparation results, Construct’s production/expression results, and Final experiment results.
In our final experiment, we used 8 different samples.
Samples 1I, 1II: E.coli surface displaying OmpA:LRR (pET-2B:OmpA:LRR) from the first colony selected
Samples 2I, 2II: E.coli surface displaying OmpA:LRR (pET-2B:OmpA:LRR) from the second colony selected
Samples NBI, NBII: E.coli surface displaying OmpA:NB-LRR (pET-2B:OmpA:NB-LRR)
Sample Ctrl: Our negative control which is E.coli DH10b cells, not expressing anything
Sample PE: Total protein extract of N. Sylvestris leaves expressing the viral Nsm115-135 protein.
Samples 1I, 1II, 2I, 2II, NBI, NBII, and Ctrl were incubated for 2 hours with the total protein extract of N. Sylvestris
After incubation time ended, we pelleted the cells, discarded the supernatant, and washed the cells with PBS (2x). At the end of the final wash, we resuspended the cells in dH2O.
In this picture, we can see the image of the way the samples were placed in the biomolecular imager.
This picture clearly shows that Fluorescence is clearly visible in all of the test samples (smaller fluorescence in sample 2I). This means that the cells bind the viral Nsm115-135 protein fused with YFP and so fluorescence is visible. A minimal fluorescent signal is visible at the negative control probably due to the low-affinity binding of some E.coli membrane protein with either YFP or Nsm115-135.
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