Briefly explain the principle behind the following techniques: (a) ion-exchange chromatography (b) gel filtration chromatography (c) affinity chromatography
Briefly explain the principle behind the following techniques: (a) ion-exchange chromatography (b) gel filtration chromatography (c) affinity chromatography
Biology: The Dynamic Science (MindTap Course List)
4th Edition
ISBN:9781305389892
Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan
Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan
Chapter19: Genomes And Proteomes
Section: Chapter Questions
Problem 12TYK
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1. Briefly explain the principle behind the following techniques:
(a) ion-exchange chromatography
(b) gel filtration chromatography (c) affinity chromatography (d) Use three diagrams to illustrate the separation of three different proteins by these methods.
2. Genetic instability in the form of point mutations, chromosome rearrangements, and epigenetic changes needs to be maximal to allow the development of cancer.
(a) With diagrams explain:
(i) point mutations
(ii) chromosome rearrangement (iii) epigenetic changes
3. Describe the process of cloning a DNA fragment into the Eco R1and Alu I sites of the vector pUC18. How would you screen for clones that contain an insert?
4. A protein has a stability ranging from 6 to 15 kcal/mole at 37 OC. Stability is a measure of the equilibrium between the folded and unfolded protein given by the relationship;
folded(F) = unfolded (U), k=[U]/[F]
For a protein with a stability of 10 kcal/mole, calculate the fraction of unfolded protein that would exist at equilibrium at 37 OC. Use the following equation: GO=RTlnk=-2.3RTlog k, where R=1.98 x 10-3 kcal/mole and T is the equilibrium constant.
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