Document Type

Thesis

Date of Award

Spring 5-17-2011

School/College

College of Science, Engineering, and Technology (COSET)

Degree Name

MS in Biology

First Advisor

Jason Rosenzweig

Abstract

In this study, the effects on transcript levels of E. coli (BC) (ATCC 4157) upon exposure to growth in space will be evaluated. EC is an enteric bacteria, thus can be found in the intestinal tracts of both healthy and diseased humans and animals. The specific transcript levels to be evaluated will be primarily based upon previous findings in which a subset of EC genes was shown to be responsive to space-like conditions, also known as Low shear modeled microgravity (Tucker et al, 2007). One such gene, accA, (which encodes acetyl coenzyme A) was found to be up-regulated in EC grown under space-like conditions. Additionally, the apt gene (which encodes adenine phosphoribosyltransferase and is involved in adenine salvage) was shown to be down-regulated EC grown under space-like conditions (Tucker et aI, 2007). Expression levels of these two genes will be evaluated in simulated microgravity grown EC samples, thereby confirming previously published ground-based simulated microgravity studies. 1 2 Simulated microgravity grown samples were grown at either 22°C for 48 hours or at 37°C for 24 hours prior to being fixed in a concentrated RNA later solution. RNA will be extracted from samples and purified using RNeasy Kit (Qiagen) and cDNA will be generated using reverse transcriptase - One-Step-RT-PCR Kit (Qiagen). The cDNA will then be subjected to quantitative, real-time PCR to determine expression level differences using QPCR Kit (Qiagen). Simulated microgravity grown EC transcript evaluation serves as a reference for future space-flight-grown EC transcript profiles which could provide insights into the responses of bonafide bacterial pathogens that could have adverse effects on the health of astronauts during space mission

Download

Share

COinS