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Modification of the mycobacteriophage Ms6 attP core allows the integration of multiple vectors into different tRNAala T-loops in slow- and fast-growing mycobacteria |
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| Authors: | Tiago Dos Vultos Isabelle Méderlé Valérie Abadie Madalena Pimentel José Moniz-Pereira Brigitte Gicquel Jean-Marc Reyrat Nathalie Winter |
| Affiliation: | 1. Unité de Génétique Mycobactérienne, Institut Pasteur, 25, rue du Dr Roux, 75724, Paris Cedex, 15, France 2. Unidade dos Retrovirus e Infec??es Associadas, Centro de Patogénese Molecular, Faculdade de Farmácia da Universidade de Lisboa, Lisbon, Portugal 3. Inserm-U 570, Unité de Pathogénie des Infections Systémiques, Groupe Avenir, Université Paris V-Descartes, Faculté de Médecine René Descartes, Paris Cedex, 15, F-75730, France |
| Abstract: | BackgroundMycobacteriophage Ms6 integrates into Mycobacterium smegmatis and M. bovis BCG chromosome at the 3' end of tRNAala genes. Homologous recombination occurs between the phage attP core and the attB site located in the T-loop. Integration-proficient vectors derived from Ms6 are useful genetic tools, but their insertion sites in the BCG chromosome remain poorly defined. The primary objective of this study was to identify Ms6 target genes in M. smegmatis and BCG. We then aimed to modify the attP site in Ms6-derived vectors, to switch integration to other tRNAala loci. This provided the basis for the development of recombinant M. bovis BCG strains expressing several reporter genes inserted into different tRNAala genes. |
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