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Team:Bielefeld-CeBiTec/Results/Biosafety/Outlook
From 2014.igem.org
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So in conclusion it could be demonstrated, that the antibiotic-free selection system by via the D-alanine auxotrophic strain DH5α <i>Δalr</i> <i>ΔdadX</i> is not only possible, but even <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosafety/TransformationEfficiency">more efficent</a> according to the transformation efficiency with the plasmid <a href="http://parts.igem.org/Part:BBa_K1465401">BBa_K1465401</a>. Addition the novel sysmtem requires a <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosfety/TransformationProcess">shorter incubation</a> in SOC-media after the successful tranformation to reach comparable transformation effiency than in Chloramphenicol. (On top it could be demonstrated that this system can be used for molecular cloning of normal plasmid size like <a href="http://parts.igem.org/Part:BBa_K1465401">BBa_I13522</a> with a total size of 4100 bp). Additionaly the selection via the complementation of the alanine racemase is suitable also for <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosafety/Long-termStability">longer time</a> and as constant as the plasmidstability with the antibiotic Chloramphenicol.<br> | So in conclusion it could be demonstrated, that the antibiotic-free selection system by via the D-alanine auxotrophic strain DH5α <i>Δalr</i> <i>ΔdadX</i> is not only possible, but even <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosafety/TransformationEfficiency">more efficent</a> according to the transformation efficiency with the plasmid <a href="http://parts.igem.org/Part:BBa_K1465401">BBa_K1465401</a>. Addition the novel sysmtem requires a <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosfety/TransformationProcess">shorter incubation</a> in SOC-media after the successful tranformation to reach comparable transformation effiency than in Chloramphenicol. (On top it could be demonstrated that this system can be used for molecular cloning of normal plasmid size like <a href="http://parts.igem.org/Part:BBa_K1465401">BBa_I13522</a> with a total size of 4100 bp). Additionaly the selection via the complementation of the alanine racemase is suitable also for <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/Biosafety/Long-termStability">longer time</a> and as constant as the plasmidstability with the antibiotic Chloramphenicol.<br> | ||
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Revision as of 19:17, 16 October 2014
Biosafety
Summary
So in conclusion it could be demonstrated, that the antibiotic-free selection system by via the D-alanine auxotrophic strain DH5α Δalr ΔdadX is not only possible, but even more efficent according to the transformation efficiency with the plasmid BBa_K1465401. Addition the novel sysmtem requires a shorter incubation in SOC-media after the successful tranformation to reach comparable transformation effiency than in Chloramphenicol. (On top it could be demonstrated that this system can be used for molecular cloning of normal plasmid size like BBa_I13522 with a total size of 4100 bp). Additionaly the selection via the complementation of the alanine racemase is suitable also for longer time and as constant as the plasmidstability with the antibiotic Chloramphenicol.
Refernces
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Kang L, Shaw AC, Xu D, Xia W, Zhang J, Deng J, Wöldike HF, Liu Y, Su J. (2011) Upregulation of MetC is essential for D-alanine-independent growth of an alr/dadX-deficient Escherichia coli strain. Journal of bacteriology, vol. 193, pp. 1098 - 1106.
