Kelly Fowlkes

Abstract

Two-dimensional gel electrophoresis (2DE) is the most commonly employed technique in the field of proteomics. The technique is designed to separate thousands proteins in highly complex biological samples. It is largely because of the diversity of the sample that 2DE has encountered problems with reproducibility. This study was designed to address and alleviate many of these problems and also to establish quantitative standards for intra-laboratory results. A universal standard gel of Pseudomonas putida KT2440 was created and tested. The universal standard gel will serve as an intra-laboratory reference to which future experiments will be compared. An acceptable coefficient of variation between future control gels and the universal standard gel was determined to be less than 35%. Another aim of this study is to explore the effects of growth time on protein expression and determine a window of time in which growth can be stopped and protein expression is preserved. Currently there have been no studies relating to this topic. Protein samples were harvested every 30 minutes throughout the log-phase of growth and their protein expression was determined. It was found that within a 30 minute window of time surrounding mid-log phase the protein expression does not change, within statistical limits. This study will serve as the quantitative cornerstone for all future intra-laboratory experiments.

Library of Congress Subject Headings

Two-dimensional electrophoresis--Standards; Pseudomonas; Proteomics

Publication Date

9-1-2005

Document Type

Thesis

Department, Program, or Center

School of Chemistry and Materials Science (COS)

Advisor

Tubbs, Laura - Chair

Advisor/Committee Member

Craig, Paul

Comments

Note: imported from RIT’s Digital Media Library running on DSpace to RIT Scholar Works. Physical copy available through RIT's The Wallace Library at: QP519.9.T84 F69 2007

Campus

RIT – Main Campus

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