Refactoring M13: Difference between revisions
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M13K07 is a helper phage vector carrying all the essential genes of for M13 phage production in addition to KanR and a p15 ori. We have | M13K07 is a helper phage vector carrying all the essential genes of for M13 phage production in addition to KanR and a p15 ori. We have created 8 mutations within the coding regions from geneII through geneIII. These mutations create new cut sites that allow for the easy insertion of synthesized DNA fragments, allowing one to quickly re-engineer the phage or insert novel elements into the genome. The mutations are shown in the figure below. The final mutant bearing all 8 mutations shows a "weakened" phenotype of about 10x below wild type cfu/ml phage titer and smaller plaques. | ||
[[Image:M13Scaffold.jpg|400 px| Entire M13K07 plasmid map showing single cutters]] | [[Image:M13Scaffold.jpg|400 px| Entire M13K07 plasmid map showing single cutters]] | ||
We are currently using this phage for instructional purposes in genome redesign in the class 20.109 [http://openwetware.org/wiki/20.109%28F07%29:Module_1]. | We are currently using this phage for instructional purposes in genome redesign in the class 20.109 [http://openwetware.org/wiki/20.109%28F07%29:Module_1]. | ||
Latest revision as of 19:57, 14 November 2007
M13K07 is a helper phage vector carrying all the essential genes of for M13 phage production in addition to KanR and a p15 ori. We have created 8 mutations within the coding regions from geneII through geneIII. These mutations create new cut sites that allow for the easy insertion of synthesized DNA fragments, allowing one to quickly re-engineer the phage or insert novel elements into the genome. The mutations are shown in the figure below. The final mutant bearing all 8 mutations shows a "weakened" phenotype of about 10x below wild type cfu/ml phage titer and smaller plaques.
We are currently using this phage for instructional purposes in genome redesign in the class 20.109 [1].
