Pearson eText for Biochemistry: Concepts and Connections -- Instant Access (Pearson+)
Pearson eText for Biochemistry: Concepts and Connections -- Instant Access (Pearson+)
2nd Edition
ISBN: 9780137533114
Author: Dean Appling, Spencer Anthony-Cahill
Publisher: PEARSON+
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Chapter 22, Problem 8P
Interpretation Introduction

Interpretation:

If a replication DNA polymerase had its 3'exonuclease site 1.5 nm from polymerase site, instead of 3.0 nm, the effect of this change in the fidelity of the enzyme should be explained. An additional change that can provide the enzyme with the same fidelity should be described.

Concept Introduction:

Polymerase is the enzyme that helps in the synthesis of nucleic acid polymers. The polymerase enzyme copies the RNA and DNA template using base-pairing techniques.

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Draw a diagram to demonstrate 3' to 5' exonuclease activity?
1. Write the piece of mRNA that would code for the amino acid sequence NH3 - met - tyr - cys- his- CO2.  Label the 5' and 3' ends. (diagram attached)  2.
You were tasked with purifying an untagged transcription factor (molecular weight 65,000Da, isoelectric point unknown) from a contaminant Protein A (molecular weight 50,000 Da, isoelectric point 5.0). You were also instructed to use the protein for crystallization studies after purification.i) In the initial purification step, which type of chromatography should you attempt? Explain your choice and specify the requirements of the buffer solution you should use. ii) Analyzing the proteins recovered from step i) using SDS-PAGE, you still observed a faint band of Protein A in addition to the transcription factor band. Given the limited time available for further purification, you must choose ONE appropriate chromatography method to maximize the chances of separating the two proteins as well as your crystallization studies. Provide a detailed explanation of your selection and the techniques/strategies you would employ to achieve this.
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