Microbiology: An Evolving Science (Fourth Edition)
Microbiology: An Evolving Science (Fourth Edition)
4th Edition
ISBN: 9780393615098
Author: John W. Foster, Joan L. Slonczewski
Publisher: W. W. Norton & Company
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Chapter 12.4, Problem 2TQ
Summary Introduction

To review:

The recovery of the gene following phage display gene and the expression as well as the purification of the protein product.

Introduction:

The technique in which DNA(deoxyribonucleic acid) sequences that encode non-phage peptides are cloned into phage capsid genes is called phage display. The synthesis of thesepeptides occur as a part of the production of the capsid protein and then are displayed on the surface of the phage. The DNA sequence can code for any product according to the requirement.Using this technique, peptide variants with altered properties can be screened.

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One of the reasons why phage therapy has not been applied widely is that bacteria can become resistant to bacteriophages as well, through mutations in genes encoding for specific proteins.  What would be a protein in the bacterial cell that, if mutated, would make that cell resistant to phage infection?
The clustered, regularly interspaced short palindromic repeats (CRISPR) system is like a bacterial adaptive immune system. After a bacteria is infected by a bacteriophage, CRISPR prevents the same type of phage from infecting the cell in the future by... integrating a phage DNA fragment into the chromosome, and transcribing it into a short RNA that binds a Cas nuclease to target and destroy invading phage DNA. modifying the attachment sites on the cell so the phage can't bind. O recognizing a molecule on the phage's surface and secreting a protein that prevents the phage from binding to the cell. recognizing a protein on the phage's surface and secreting an enzyme that destroys the phage.
Ahmed had isolated bacteria from the sputum of a patient that seemed somewhat resistant to their antibiotic treatment, since multiple weeks of treatment had yielded no satisfactory improvement of their active disease. The researcher had then resuspended the patient's sputum in saline and used a mouse model to measure the efficiency of a phage selected for its capacity to target M. mycobacterium, as a single therapy, and as a combination therapy with antibiotics. Here is a schematic representation of the experiment: Group 1 Control (saline) Inoculate mice with patient sputum Group 2 wait for active Treated with phage disease to show Collect lung sample every 2 weeks Tuberculosis Sputum samples patient Group 3 Treated with Abx Group 4 Split mice showing active disease in 4 Treated with Abx and Count bacteria in sample groups phage Figure 4. Sputum from patients were isolated and resuspended in saline. Mice were given equal amounts of resuspended sputum intra-nasally and were left to…
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