کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2824526 | 1161665 | 2006 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Allelic replacement in Staphylococcus aureus with inducible counter-selection Allelic replacement in Staphylococcus aureus with inducible counter-selection](/preview/png/2824526.png)
A method for rapid selection of allelic replacement mutations in the chromosome of Staphylococcus aureus is described. Plasmid pKOR1, an Escherichia coli/S. aureus shuttle vector, permits rapid cloning via lambda recombination and ccdB selection. Plasmid transformation of staphylococci and growth at 43 °C, a non-permissive condition for pKOR1 replication, selects for homologous recombination and pKOR1 integration into the bacterial chromosome. Anhydrotetracycline-mediated induction of pKOR1-encoded secY antisense transcripts via the Pxyl/tetO promoter, a condition that is not compatible with staphylococcal growth, selects for chromosomal excision and loss of plasmid. Using this strategy, allelic replacements in S. aureus rocA were generated at frequencies that obviated the need for antibiotic marker selection.
Journal: Plasmid - Volume 55, Issue 1, January 2006, Pages 58–63