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Genetic manipulation of Streptococcus pyogenes (Group A Streptococcus, GAS) has historically been a challenging process, with considerable variation in efficiency between different strains. Here, we outline an optimized, rapid method for creating markerless isogenic mutations that combines Gibson assembly cloning with a new temperature-sensitive plasmid, pLZts. This method is highly efficient and reduces the time needed to create GAS mutants to ~2-3 weeks, with the ability to prepare multiple mutants simultaneously.
|Title of host publication||Group A Streptococcus|
|Subtitle of host publication||Methods and Protocols|
|Editors||Thomas Proft, Jacelyn M. S. Loh|
|Place of Publication||USA|
|Publisher||Humana Press Inc.|
|Number of pages||11|
|Publication status||Published - 2020|
|Name||Methods in Molecular Biology|
FingerprintDive into the research topics of 'Genetic Manipulation of Group A Streptococcus-Gene Deletion by Allelic Replacement'. Together they form a unique fingerprint.
- 1 Finished
Currie, B., Carapetis, J., Anstey, N., McBride, W., Chang, A., Garvey, G., Maple-Brown, L. & Sorrell, T.
1/01/17 → 31/12/21