Bacterial transformation involves inserting a gene from one organism into another using plasmids. Plasmids containing a gene of interest are introduced into bacteria, allowing the bacteria to express proteins from that gene. In this workshop, green fluorescent protein (GFP) was used as a marker to test if E. coli bacteria were transformed. E. coli cultures were grown, one containing a GFP plasmid. Under UV light, the plasmid-containing culture showed GFP expression, indicating transformation occurred. SDS-PAGE was used to separate and observe the transformed bacteria's proteins, revealing bands corresponding to GFP.

1. Workshop: pGLO Bacterial Transformation
By Belinda Román and Ricardo Chiesa
Bacterial transformation involves the process of inserting a certain gene from one
organism into another organism. The purpose of this is to make an organism express a gene that
is not present in its own genome. Scientists use different techniques to do so, and one of them is
the use of a plasmid. Plasmids are pieces of an autonomous replicating DNA obtained from
bacteria. Genes of interest are inserted into plasmids, and then by introducing plasmids into
bacteria, transformation will occur As a result, the host organisms will express the proteins of
interest. In the lab, we worked with this process, but we used Green Fluorescent Protein (GFP) as
a biological marker to see if bacterial transformation occurs. First of all, we make cultures of E.
coli bacteria in Petri dishes. One of the cultures had the plasmid that was going to transform the
bacteria. The following week, we looked at this culture with UV light, and transformation did
occur because GFP was expressed in E. coli. Afterwards, we practiced the technique of SDS-PAGE
to divide and observe proteins of the transformed bacteria in a polyacrylamide gel. Finally, we
observe the proteins with UV light and Coomassie staining to see the location of GFP. Our results
show that partially denatured proteins present a band of 37kD, and the completely denatured
proteins show a band of 27kD.