Protein Purification: Isolating Fluorescent Protein from Bacteria
Protein Purification Lab Introduction
This lab follows either “Bacterial Transformation” or “Bacterial Transformation with Gene Regulation.”
Obtaining purified protein from a genetically modified organism (GMO) is an important step for many pharmaceutical companies and research labs. Whether manufacturing proteins for clinical use, such as insulin, or studying the function of proteins in biochemical assays, protein purification is an essential process. Read More
In this lab, students will use nickel affinity chromatography to isolate fluorescent proteins from E. coli bacteria that they have previously transformed with the pKiwi plasmid (or another BABEC plasmid).
Nickel affinity chromatography utilizes six histidine (his) amino acids that have been added to the recombinant green fluorescent protein as an artificial “tag.” This 6XHis tag has a high affinity for nickel resin. Since only the recombinant protein will possess this unique tag, the purification process can specifically separate FPs from the other proteins in the cell lysate.
This lab provides an opportunity for hands on exploration of the applications of genetic engineering. It can introduce or review topics ranging from recombinant DNA, transcription, translation, protein expression, cell lysis and clearly demonstrates how proteins can be isolated for use in research or medicine.
Lesson Resources Student Guide – Download and edit student worksheets to be completed during the Protein Purification Lab. Lesson Slides – Articulates with the student guide above and accompanying lesson plans below. Lesson Plan – Suggested times for pacing, sequence and activities for the lab. Includes vocabulary tool and other lesson resources. Supplemental Resources – Includes background information, outside articles, videos, and more related to the lab