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- W2091156802 abstract "Insertional mutagenesis is a technique often used to inactivate genes in Streptococcus pneumoniae. Using conventional vectors, a 5' segment of the targeted gene remains under the control of the gene's authentic promoter following gene disruption. Thus, the expression of a functional peptide and the misinterpretation of results in consequence cannot be excluded. To circumvent this problem, we have developed a plasmid for insertional mutagenesis based on the tmRNA-tagging system of S. pneumoniae which ensures that any protein expressed after gene disruption is degraded. Insertional mutagenesis using this vector results in the targeted gene being tagged with a tmRNA-derived sequence coding for a proteolysis tag. Here we show that the translation product of a gene tagged by this method is not detectable by Western blotting, suggesting that the protein was degraded. This modified vector allows total inactivation of genes with a reliability that cannot be achieved by conventional vectors for insertional mutagenesis. This approach can be applied to other bacterial species." @default.
- W2091156802 created "2016-06-24" @default.
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- W2091156802 date "2000-10-01" @default.
- W2091156802 modified "2023-09-25" @default.
- W2091156802 title "An improved vector system for insertional gene inactivation inspired by the tmRNA-tagging system of S. pneumoniae" @default.
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- W2091156802 doi "https://doi.org/10.1016/s0167-7012(00)00173-1" @default.
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