Title Information
Title
Stability and location of mutant proteins in bacterial cells
Name: Personal
Name Part
Spangle, Dylan
Role
Role Term: Text
creator
Name: Personal
Name Part
Weinreich, Daniel
Role
Role Term: Text
advisor
affiliation
Brown University. Department of Ecology and Evolutionary Biology
Name: Corporate
Name Part
Brown University. Undergraduate Teaching and Research Award
Role
Role Term: Text
research program
Type of Resource
still image
Genre (aat)
posters
Origin Information
Place
Place Term: Text
Providence
Publisher
Brown University
Date Created (encoding="w3cdtf")
2015-08-07
Physical Description
Extent
1 poster
digitalOrigin
reformatted digital
Abstract
Evolution by selection begins at the molecular level within the cell: a series of mutations occur that are able to become fixed in the population, and finally lead to a protein sequence that performs a novel function for the organism. ß-lactamase, a bacterial protein that counteracts ß-lactam antibiotics such as ampicillin, is a significant and well-studied protein, making it an excellent model for studying the biochemical pathway of protein evolution. It has been hypothesized that the relative fitness of mutant proteins is primarily determined by three factors: catalytic activity at the active site, stability of overall folding structure, and localization in the cell. We expressed and purified wild-type and mutant alleles of the TEM-1 ß-lactamase gene to study stability and localization of this protein. All of these mutations were outside the protein's active site, removing catalytic activity at the active site as a variable. We isolated proteins from periplasmic, cytosol-soluble and insoluble fractions using established biochemical methods. Subsequently these protein samples be analyzed using transverse urea gradient gel electrophoresis and Western blot to determine stability of these alleles, and using ELISA to quantitatively determine localization.
Subject (LCSH)
Topic
Drug resistance in microorganisms
Subject (LCSH)
Topic
Beta-lactamases
Subject (LCSH)
Topic
Biochemistry
Identifier: DOI
10.26300/a40q-2727