Document Type
Article
Publication Date
5-11-2012
Publication Title
Journal of Biological Engineering
Department
Thayer School of Engineering
Abstract
Temperature-sensitive (Ts) plasmids are useful tools for genetic engineering, but there are currently none compatible with the gram positive, thermophilic, obligate anaerobe, Clostridium thermocellum. Traditional mutagenesis techniques yield Ts mutants at a low frequency, and therefore requires the development of high-throughput screening protocols, which are also not available for this organism. Recently there has been progress in the development of computer algorithms which can predict Ts mutations. Most plasmids currently used for genetic modification of C. thermocellum are based on the replicon of plasmid pNW33N, which replicates using the RepB replication protein. To address this problem, we set out to create a Ts plasmid by mutating the gene coding for the RepB replication protein using an algorithm designed by Varadarajan et al. (1996) for predicting Ts mutants based on the amino-acid sequence of the protein.
DOI
10.1186/1754-1611-6-5
Original Citation
Olson DG, Lynd LR. Computational design and characterization of a temperature-sensitive plasmid replicon for gram positive thermophiles. J Biol Eng. 2012 May 11;6(1):5. doi: 10.1186/1754-1611-6-5. PMID: 22578246; PMCID: PMC3464808.
Dartmouth Digital Commons Citation
Olson, Daniel G. and Lynd, Lee R., "Computational Design and Characterization of a Temperature-Sensitive Plasmid Replicon for Gram Positive Thermophiles" (2012). Dartmouth Scholarship. 1241.
https://digitalcommons.dartmouth.edu/facoa/1241
