|
|
|
CH 150: Introduction to Biochemistry pGLO: Bacterial Transformation Bacterial transformation occurs when a bacterial cell takes up and expresses new DNA. Usually this new DNA is added as a plasmid, a small circular piece of DNA. This techniques is very powerful, in that we can thus insert a gene for a specific protein, for example, into a plasmid by use in restriction enzymes. This newly engineered plasmid is then introduced into the bacteria, which then express the protein gene product of interest. In this experiment, we will be transforming a colony of E. coli with a plasmid containing the gene called pGLO. This gene encodes the green fluorescent protein from the bioluminescent jellyfish, Aequorea victoria. You will select a colony of normal bacteria and perform the transformation during the Friday lab. We will need to meet for a VERY short period (30 minutes at the moist) on Saturday to select transformed colonies for the next experiment, Protein Purification, where you will isolate the green fluorescent protein now produced by your new clones. We will be using a kit developed by Bio-Rad Laboratories as a part of their Biotechnology Explorer Curriculum. Follow the link to the Student Manual. Print a copy of the manual to follow in the lab. Your completed copy of the manual will serve as your lab report. |