We successfully produced recombinant plasmid, with 6.4 KBP (6389 DNA base pairs). By the process of transformation, we have successfully bioengineered a kind of bacteria, which can produce the protein that we want. The following picture indicates that the incubated bacteria, which was on an agar plate and it was stored for further usage. We intentionally added antibiotics in the agar plate to test whether the bacteria can be incubated successfully. If it can’t be killed by those specific antibiotics, it means that the bacteria is the specific bacteria with our recombinant plasmid.
To prove that the modified bacteria has our designed gene, we used gel electrophoresis to check whether the bacteria contains our gene or not. In the picture below, we can see the band of our bacteria at the position around 6.4 KBP of the ladder, which is exactly the length of our gene. This indicates that the bacteria contains our designed gene.
To prove that the modified bacteria has GFP (green fluorescent protein), we can see the green fluorescence in the incubated BL21 bacteria under a blue light source in the picture below.
(IPTG is used to induce the expression of the gene in BL21)
Since this solution shows fluorescence, we extracted the protein by cell lysis and underwent protein purification. To prove that the bacteria can produce the protein we want, we test the extract by SDS-page.
The band in the range 34 kDa proves the expression of TP1_P5_GFP
Since the protein produced in BL21 matches the estimated kDa of our fusion protein, the plasmid insertion is proved to be successful, and our desired protein can be produced.
The band in the range 34 kDa proves the expression of TP1_P5_GFP
Since the protein produced in BL21 matches the estimated kDa of our fusion protein, the plasmid insertion is proved to be successful, and our desired protein