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Site‐specific integration of the Streptomyces plasmid pSAM2 in Mycobacterium smegmatis
Author(s) -
Martin C.,
Mazodier P.,
Mediola M. V.,
Gicquel B.,
Smokvina T.,
Thompson C. J.,
Davies J.
Publication year - 1991
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1991.tb02095.x
Subject(s) - mycobacterium smegmatis , biology , plasmid , microbiology and biotechnology , mycobacterium , streptomyces , computational biology , genetics , bacteria , mycobacterium tuberculosis , dna , tuberculosis , medicine , pathology
Summary A method which allowed the stable integration of DNA fragments at a single site ( attB ) in the chromosome of Mycobacterium smegmatis was developed using an integrative element from Streptomyces ambofaciens , pSAM2. Vectors containing an Escherichia coli replicon (pBR322), the kanamycin resistance gene from Tn 903 for selection in mycobacteria, and a fragment of pSAM2 containing the int gene as well as the attachment site ( attP ) were constructed and introduced to M. smegmatis by electroporation. Transformants showed stable integration of the plasmid into a single site ( attB ) of the mycobacterial genome. This approach should be valuable for analyses of gene expression in various mycobacterial species and permit the development of stable recombinant mycobacterial vaccine strains expressing bacterial or viral genes inserted in pSAM2.