BIOL 2070- Midterm Exam Guide - Comprehensive Notes for the exam ( 14 pages long!)
Document Summary
Scheich, c. , sievert, v. , and b ssow, k. 2003. An automated method for high- throughput protein purification applied to a comparison of his-tag and gst-tag affinity chromatography. The authors inform the reader of the background of protein and how it assists in discovering how a gene functions. They further explain how to use the protein by purifying it free from other unwanted substances using affinity chromatography due to its efficiency. Escherichia coli can be used to overexpress the proteins by crystallization. For purifying the proteins, two affinity tags; hexahistidine tag (his-tag) or glutathione s- transferase (gst)-tag can be used from the e. coli expression system. The his-tag tagged to nickel ions, whereas gst-fusion proteins were purified using glutathione affinity chromatography. The expressed proteins were bounded with the beeds through incubation. To separate the beads with other substances, filter plates were used which only retained the beads. After purifying the selected proteins, it was concluded that the.
