From: I-TRAP: A method to identify transcriptional regulator activated promoters
Strains | Relevant Characteristics | Source/Reference |
---|---|---|
Electromax DH5α | Cloning host | Invitrogen |
M. smegmatis LR222 | Cloning host | [20] |
M. smegmatis::SEint.2 | constitutively expresses M. tuberculosis sigE gene | This study |
M. smegmatisΔsigE | sigE knockout mutant | This study |
M. tuberculosis H37Rv TMC102 | Virulent laboratory strain | [21] |
Plasmids | Â | Â |
pAWS2 | Source of the lacZ gene | R. Van Landingham |
pBluescript-int | Source of integrase gene, int | B. Plikaytis |
pCL5 | Backbone for pCL8 | [9] |
pCL8 | Source of the rpsL promoter | This study |
pCRScript SK (+) | Vector with multiple cloning site | Stratagene |
pMV261 | Source of the hsp60 promoter and OriM | [22] |
pPICZalphaC | Source of bleomycin resistance gene, ble | Invitrogen |
pPR27 | Source of gentamicin resistance gene, aacC1 | [23] |
pSE.1 | pUC19 with promoterless aph and xylE genes | This study |
pSE.2 | pSE.1 with the hyg gene | This study |
pSE.3 | pSE.2 with the OriM | This study |
pSE.4 | promoter-trap vector | This study |
pSEint.1 | Integrating plasmid containing the ble gene | This study |
pSEint.2 | Integrating plasmid, constitutive M. tuberculosis sigE expression | This study |
pSEKO.1 | 5' region of the M. smegmatis sigE gene | This study |
pSEKO.2 | pSEKO.1 with 3' end of M. smegmatis sigE gene | This study |
pSEKO.3 | pSEKO.2 with the aacC1 gene | This study |
pSEKO.4 | Suicide plasmid with interrupted M. smegmatis sigE gene | This study |
pT7Blue | Blunt cloning vector | This study |
pTKMX97 | Source of the xylE gene | [10] |
pUC19 | Vector with multiple cloning site | Amersham Biosciences |
pUC4-KIXX | Source of aph gene | Amersham Biosciences |
pUS267 | Source of hygromycin resistance gene, hyg | J. Dale |