Team:GeorgiaTech/Abstract

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Revision as of 03:42, 29 September 2011

Title: De Novo Adaptation of Streptococcus thermophilus CRISPR1 Defense in E. coli

Abstract:
The ultimate goal of our project was to utilize this system as a mobile gene targeting system capable of eliminating antibiotic resistance. The protoype construct, pSTINGER, was designed for quantitative modeling, proof of concept, and vaccination on a single plasmid. As a result of experimental error, intellectual property agreements, material availability, our biobrick constructs remain experimentally theoretical, though we have modeled our system in various scenarios.

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-A diverse range of Bacteria and Archaea acquire resistance to foreign DNA by integrating short fragments of the invading nucleic acid into clusters of regularly interspaced short palindromic repeats (CRISPRs) on their genomic DNA. For our project we have PCR amplified the CRISPR1 locus from the chromosome of Streptococcus thermophilus [LMD-9] and ligated it into an integration vector to place it on the chromosome of Bacillus subtilis through allelic recombination on the chromosome. B. subtilis served as our model organism because it does not naturally posses a CRISPR mechanism. This should demostrate that the S. thermophilus CRISPR1/Cas system can be transferred into Bacillus subtilis and provide heterologous protection against plasmid transformation and phage infection.
+The ultimate goal of our project was to utilize this system as a mobile gene targeting system capable of eliminating antibiotic resistance. The protoype construct, pSTINGER, was designed for quantitative modeling, proof of concept, and vaccination on a single plasmid. As a result of experimental error, intellectual property agreements, material availability, our biobrick constructs remain experimentally theoretical, though we have modeled our system in various scenarios.
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