Abstract:
:Gene disruption experiments play an important role in the functional characterization of genes in mycobacteria and rely mostly on the use of one or two antibiotic resistance markers. We have developed a system for mycobacteria which features both the advantages of the use of antibiotic resistance markers for gene disruption experiments and the ability to efficiently rescue the marker leaving an unmarked mutation on the chromosome. This new genetic tool relies on the transposon gammadelta site-specific recombination system. A res-OmegaKm-res cassette was used to generate an insertional mutation by allelic exchange both in Mycobacterium smegmatis and Mycobacterium bovis BCG. Upon expression in the mutated strains of tnpR, the transposon gammadelta resolvase gene, res-OmegaKm-res, was excised efficiently leaving behind a single res sequence at the mutated locus. A plasmid was engineered allowing expression of tnpR from an easily curable mycobacterial vector. This system will be useful for simple construction of unmarked mutations or repeated use of the same antibiotic marker to generate multiple mutants.
journal_name
FEMS Microbiol Lettjournal_title
FEMS microbiology lettersauthors
Malaga W,Perez E,Guilhot Cdoi
10.1016/S0378-1097(03)00003-Xkeywords:
subject
Has Abstractpub_date
2003-02-28 00:00:00pages
261-8issue
2eissn
0378-1097issn
1574-6968pii
S037810970300003Xjournal_volume
219pub_type
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
pub_type: 杂志文章
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journal_title:FEMS microbiology letters
pub_type: 信件
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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journal_title:FEMS microbiology letters
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