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Team:Glendale CC AZ/Project/ExperimentalOverview
From 2013.igem.org
Experimental Overview
We began our project with a plan to characterize existing biobricks, as well as create a few of our own to complete a comprehensive desiccation toolkit that can be used for a variety of applications. Osaka’s 2011 iGEM team created coding device biobricks for genes PprI (BBa_K602000), PprM (BBa_K602002), PprA (BBa_K602001), and RecA (BBa_K602003) as well as expression parts for PprI (BBa_K602005), PprM (BBa_K602007), and PprA (BBa_K602006). We wanted to characterize these parts in a novel way, as Osaka did not expose their transformed bacteria to salt stress. We also planned to expose the transformed bacteria to our Hydrogen Peroxide Growth Curve Assay for further characterization. University College London’s 2012 iGEM team created a coding part for IrrE (BBa_K729001), as well as an expression device containing their coding sequence for IrrE (BBa_K729005). We planned to further characterize their IrrE biobricks by exposing the transformed E.coli to our Salt Stress Growth Curve Assay as well as our Hydrogen Peroxide Growth Curve Assay. Valencia’s 2010 team created an expression part for one of the LEA proteins, which also is said to confer salt and oxidative resistance when transformed in E.coli. Our goal was to further characterize their LEA biobrick. Through rigorous research using bioinformatics, we found that homologs to these genes exist in a local bacterium, D. hopiensis. Our plan was to create coding devices and expression devices for PprI, PprM, PprA, RecA, IrrE, and LEA from the local D. hopiensis bacterium. We wished to transform these genes from D. hopiensis to E.coli, as E.coli is a well-known, easily accessible, and commonly used chassis. We, would then expose the transformed bacteria to salt stress and hydrogen peroxide stress, and carefully characterize the effects.
