Fig. 1
From: Site-specific bacterial chromosome engineering mediated by IntA integrase from Rhizobium etli
Diagram of the IntA site-specific integration approach. a A novel region (the Landing Pad or LP sector) was inserted into the chromosome of R. etli. This region comprises a new attA region, flanked by a lac promoter and a promoterless green fluorescent protein (GFP) gene; and a spectinomycin resistance gene with its own promoter. b Mobilizable kanamycin-resistant donor vector (pK18 mob att Δplac) containing an attD site and a Multi Cloning site (MCS). The plac promoter was removed from the donor vector as described in Methods. c Predicted structure of integrants of pK18 mob att Δplac into the LP sector. Note that integration of the donor vector by attA X attD recombination abolishes transcription of the GFP gene, leading to nonfluorescent colonies. In panels a and c, the location of oligonucleotide primers useful to verify insertion, are indicated as arrows below the appropriate locations