pGLO Lab

In class today, we made e coli turn fluorescent green in the dark! We put a starter e coli and transformation solution into 4 micro centrifuges. We put the pGLO plasmid into only two of them. We heat shocked the centrifuges so that the bacteria would absorb the plasmid. We finally put them onto agar and incubated them for 48 hours. The results were very interesting

Plate	Number of Colonies	Color of colonies under room light	Color of colonies under UV light
-pGLO LB	carpet	grey	grey
-pGLO LB/amp	none	none	none
+pGLO LB/amp	104	grey	grey
+pGLO LB/amp/ara	64 + some carpet	grey	neon green

Our transformed bacteria glowed neon green and had resistance to ampecillin,an antibiotic.

We spread 100 microliters of bacteria onto each agar plate. In the 100 microliters, there was about 90 e coli in the solution E coli is about 1 to 2 microliters in length. It is very packed in, so the number must be closer to 100.

In the one plate of bacteria that glowed, we put arabinose. The arabinose sugar activated the pGLO gene and allowed for it to be expressed.

There are many uses for this technology. A few examples include studying cancer in mice, removing the malaria gene from mosquitoes, and studying the spread of HIV. Cancer cells can be easily watched when they have the Green Fluorescent Protein. Scientists created a gene that destroyed the GFP gene in a mosquitoes' testicles. The next step is to create a gene that will destroy the gene critical for malaria transmission. Researchers were able to see how HIV spreads from one cell to another by placing the GFP gene in an infected T-cell.

pGLO is only one application of genetic engineering. Another is crop resistance to parasites. If one variation of corn is resistant to a parasite, it is more likely to survive. Farmers genetically engineer their corn to make it resistant to certain parasites.

Our plates