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Table 1 Plasmids and strains used in this study

From: Enhanced oxygen consumption in Herbaspirillum seropedicae fnr mutants leads to increased NifA mediated transcriptional activation

Plasmids Relevant characteristic Source
pLAFR3.18 pLAFR vector containing the polycloning site of pTZ18R, TcR [8]
pRAMM1 H.seropedicae NifA expressed from lac promoter [26]
pK18mobsacBKm Allelic exchange suicide vector; mobilized by E. coli S17-1 λpir, sacB,KmR [22]
pJQ200SK Allelic exchange suicide vector; mobilized by E. coli S17-1, sacB,GmR [23]
pRAM1T7 H.seropedicae NifA in pT7-7 vector This study
pRAM1T7del H.seropedicae NifA with a deletion of 576 bp This study
pK18nifAdel XbaI/BamHI fragment from pRAM1T7del This study
pUC57Simple-3xFlag 5′-BamHI, KpnI, XhoI – 3xFlag-Stop – HindIII, SalI, XmaI-3′, AmpR Genscript® Corporation
pUC57nifAFlag H. seropedicae nifA-3xFlag gene plus 647 bp of the downstream region, AmpR This study
pK18nifAFlag H. seropedicae nifA-3xFlag gene plus 647 bp of the downstream region, Mob, SacB, KmR This study
pUC57fnr1Flag H. seropedicae fnr1-3xFlag gene plus 1000 bp of the downstream region, AmpR This study
pJQfnr1Flag H. seropedicae fnr1-3xFlag gene plus 1000 bp of the downstream region, Mob, SacB, GmR This study
pPW452 lacZ fusion vector, TcR, Mob [27]
pEMS140 TcR, Mob, nifB::lacZ fusion, nifB promoter cloned in pPW452 [6]
pRW1 TcR, Mob, nifA::lacZ fusion, nifA promoter cloned in pMP220 [20]
pRWC pRW1, but with a 49 bp deletion including the UAS 2 site for NifA [20]
pRW3 pRW1, but with a double mutation at the UAS 2 site for NifA (TGT ->TCT and ACA -> AGA) [20]
pRW22A pRW1, but with a single mutation at the −24/-12 promoter (G -> T at −25 residues) [20]
Strains Relevant characteristic Source
SmR1 Herbaspirillum seropedicae Z78 but SmR 100 μg/mL, Nif+ [24]
MB1 Derived from SmR1 fnr1 deletion [18]
MB2 Derived from SmR1, fnr2 deletion [18]
MB3 Derived from SmR1, fnr3 deletion [18]
MB13 Derived from MB1, fnr1 and fnr3 double deletion [18]
MB21 Derived from MB2, fnr1 and fnr1 double deletion [18]
MB23 Derived from MB2, fnr2 and fnr3 double deletion [18]
MB231 Derived from MB23, fnr1, fnr2 and fnr3 triple deletion [18]
MBN1 Derived from SmR1, but with 576 bp deletion in the nifA gene This study
MBN2 Double fnr1, fnr3 deletion, plus a 576 bp deletion in the nifA gene This study
MBN3 Triple fnr deletion, plus a 576 bp deletion in the nifA gene This study
MBN4 Derived from SmR1, but with a C-terminal 3xFlag nifA gene This study
MBN5 Double fnr1, fnr3 deletion, plus a C-terminal 3xFlag nifA gene This study
MBN6 Triple fnr deletion, plus a C-terminal 3xFlag nifA gene This study
MBF1 Derived from SmR1, but with a C-terminal 3xFlag fnr1 gene This study