Table 1 Strains, plasmids, and primers used in this study
Strains, plasmids, and primers | Significant properties | Source or purpose |
|---|---|---|
Strains | ||
 E. coli DH5α | General cloning host | Transgen Biotech |
 Mycobacterium neoaurum TCCC 11,978 (MNR) | Wild type | Tianjin University of Science and Technology Culture Collection Center (TCCC) |
 MNR-prpR | prpR overexpressed strain of MNR | This work |
 MNR-prpDBC | prpDBC overexpressed strain of MNR | This work |
 MNRΔglnR | Deletion of glnR in MNR | This work |
 MNRΔglnR::glnR | glnR complemented in MNRΔglnR | This work |
 MNR-prpR/ΔglnR | prpR over-expressed strain of MNRΔglnR | This work |
 MNR-prpDBC/ΔglnR | prpDBC over-expressed strain of MNRΔglnR | This work |
 MNR-pMV261 | MNR carrying vector pMV261 | This work |
Plasmids | ||
 pMV261 | Shuttle vector of mycobacterium and E. coli, carrying the heat shock hsp60 promoter, KanR | Dr. W. R. Jacobs Jr |
 p2NIL | Plasmid for allelic exchange, non-replicative in Mycobacterium species, KanR | Dr. T. Parish |
 pGOAL19 | lacZ, hyg and sacB marker genes cassette-containing vector, HygR | Dr. T. Parish |
 pMV261-prpR | pMV261 carrying an extra prpR for overexpression, KanR | This work |
 pMV261-prpDBC | pMV261 carrying an extra prpDBC for overexpression, KanR | This work |
 pMV261-glnR | pMV261 carrying an extra glnR for overexpression, KanR | This work |
 pKO-glnR | p2NILcarrying the homologous arms of glnR and the selection markers from pGOAL19 | This work |
Primers | ||
 PCR for overexpression | ||
   prpR-f | GGATCCAGCTGCAGAATTCGTGGCGAAGACATTCGCGGG | prpR amplification |
   prpR-r | CGCTAGTTAACTACGTCGACTCAGTTGGCGGGCGGGTAGG | prpR amplification |
   prpDBC-f | GGATCCAGCTGCAGAATTCGTGCGGATCATGCAGAAACA | prpDBC amplification |
   prpDBC-r | CGCTAGTTAACTACGTCGACTCACCGGCGACGTTCCAGCG | prpDBC amplification |
 PCR for deletion | ||
   glnR-U-f | ATAAACTACCGCATTAAAGCTTTCTGAACCCGTCCAGGTCGA | glnR deletion |
   glnR-U-R | CTCGGGACCGACGATCGCGATATCGGCG | glnR deletion |
   glnR-D-f | ATCGTCGGTCCCGAGTACGAGGCGCTG | glnR deletion |
   glnR-D-R | TGACACTATAGAATACATAGGATCCGGTCCGCGGACGGAAAGTGA | glnR deletion |
 For qRT-PCR | ||
   16 s-f-RT/16 s-r-RT | GTAGGGTCCGAGCGTTGTC/GCGTCAGTTACTGCCCAGAG | Quantitative RT-PCR |
   prpR-f-RT/prpR-r-RT | CCTGGGAACCCCGAAGA/GCCCGACATAGGGAAACG | Quantitative RT-PCR |
   prpD-f-RT/prpD-r-RT | TGGTCAAGGGCATCTGTCTG/TGCCCGATGGCGTTGTA | Quantitative RT-PCR |
   prpB-f-RT/prpB-r-RT | GACCAGGTGAACCCCAAGC/GCGGGCACAGATCACAAAG | Quantitative RT-PCR |
   prpC-f-RT/prpC-r-RT | GCCCAATCTGGATTTCCC/AGCGTTTGAAGCGGTTTG | Quantitative RT-PCR |
   glnR-f-RT/glnR-r-RT | CCTCGGGGAGTTGGTCAT/ GTTGGGCCAGATACTTCAGC | Quantitative RT-PCR |