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AssigneeLaboratorio Medinfar Produtos Farmaceuticos, LDA.
InventorsJose A. Frazao Moniz Pereira, Alcino Freitas Vieira, Elsa M. Ribeiro Dos Santos-Anes, Miguel A. Da Costa Garcia, Paulo J. Da Silva Alves
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The present invention discloses a DNA fragment that can direct the insertion of heterologous DNA into a specific site (tRNAAla gene) of Mycobacterium spp. genome. This DNA fragment includes the attachment site region (attP) and the integrase gene of the mycobacteriophage Ms6. Heterologous DNA linked to this DNA fragment can be carried into the mycobacterial genome through a site-specific integration mechanism. A new DNA integration process is disclosed that consists in providing the integrase gene on a suicide vector, that transiently produces the integrase required for the insertion of the gene or genes linked to the Ms6 attP region contained in a separated plasmid vector. This process allows the construction of highly stable recombinant mycobacteria which may be used as vaccines and/or therapeutic vehicles.
A genetic system for integrating DNA into a specific site of a Mycobacterium spp. genome comprising:
A purified and isolated DNA sequence of the integration locus from mycobacteriophage Ms6 DNA consisting of the nucleotide sequence shown in SEQ ID NO:16.
A genetic system for integrating DNA into a specific site of a Mycobacterium spp. genome comprising:
A purified and isolated DNA from mycobacteriophage Ms6 that promotes the transcription of DNA in Mycobacterium spp. consisting of the nucleotide sequence shown in SEQ ID NO:52.
An isolated DNA sequence that hybridizes under stringent conditions with a sequence consisting of SEQ ID NO:
An isolated amino acid sequence shown in SEQ ID NO:17.
A genetic system for integrating DNA into a specific site of a Mycobacterium spp. genome consisting of:
Dependencies are inferred from the claim text.