Targets for pSAM2 integrase-mediated site-specific integration in the Mycobacterium smegmatis chromosome.
An improved integrative cassette from plasmid pSAM2 has been constructed containing plasmid int and attP genes but excluding the xis gene, which should results in increased stability by suppression of the excision reaction. This cassette was included in both suicide and thermosensitive plasmids and used for integration in Mycobacterium smegmatis. Suicide plasmids containing this cassette integrated at a single site (attB1) in the M. smegmatis chromosome. The sequence of the attB1 site has been determined and was identified as a putative tRNA(Pro) gene. Thermosensitive plasmids containing the cassette integrated both at the same attB1 site and at other different sites, often giving rise to simultaneous integration at two sites. A second integration site (attB2) has been sequenced, which was located in the region encoding 16S rRNA of one of the two rrn operons of M. smegmatis.
Seoane
Asunción
A
Departamento de Biología Molecular, Facultad de Medicina, Universidad de Cantabria, Cardenal Herrera Oria s/n, 39011 Santander, Spain.
Navas
Jesús
J
Departamento de Biología Molecular, Facultad de Medicina, Universidad de Cantabria, Cardenal Herrera Oria s/n, 39011 Santander, Spain.
García Lobo
Juan M
JM
Departamento de Biología Molecular, Facultad de Medicina, Universidad de Cantabria, Cardenal Herrera Oria s/n, 39011 Santander, Spain.
D001483:Base Sequence; D002874:Chromosome Mapping; D002876:Chromosomes, Bacterial / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D019426:Integrases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000201:enzymology / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D009876:Operon; D010957:Plasmids / Q000235:genetics*; D012361:RNA, Transfer, Pro / Q000235:genetics; D011995:Recombination, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial; D012361:RNA, Transfer, Pro; D019426:Integrases