Transformation of Escherichia coli with the pGLO Plasmid: Going beyond the Kit

Abstract

The Bio-Rad pGLO bacterial transformation kit is commonly used to demonstrate this form of genetic exchange, which occurs in bacteria and eukaryotes and which differs fundamentally from transduction and conjugation. The basic experiment leads to the formation of green fluorescent colonies of Escherichia coli and can be extended to illustrate the specificity of the interaction between sugars and the AraC protein, the phenomenon of carbon catabolite repression, the substrate specificity of the β-lactamase encoded by the plasmid, and the role of host restriction/ modification systems in the transformation process. pGLO DNA also can be isolated using plasmid mini-prep kits, analyzed with restriction endonucleases, and used to study the conditions for transformation in more detail.