Figure 4From: Enhanced oxygen consumption in Herbaspirillum seropedicae fnr mutants leads to increased NifA mediated transcriptional activationThe NifA protein levels are higher in H. seropedicae strains lacking both Fnr1 and Fnr3. Protein extracts from different H. seropedicae strains were prepared and then hybridized with ANTI-FLAG antibodies as described in Methods. (A) Western hybridization of protein samples after resolution on 12% SDS-PAGE and transfer to PVDF membrane. (B) Protein loading control SDS-PAGE of samples used for hybridization in A. Lanes are as follows: 1, Protein molecular weight standard (MW); 2, H. seropedicae MBF1 (fnr1-3xFlag); 3, H. seropedicae MBN4 (nifA-3xFlag); 4, H. seropedicae MBN5 (nifA-3xFlag in the double fnr1, fnr3 deletion background); 5, H. seropedicae MBN6 (nifA-3XFlag in the triple fnr mutant background); 6, H. seropedicae SmR1 (no Flag protein control). In A, the MW used was Precision Plus Protein WesternC™ Pack (BioRad #161-0385), while in B, the MW used was the LMW-SDS Marker Kit (GE healthcare# 17-0446-01). Thin arrows indicate the molecular weight in KDa of protein standards. The black thick arrow indicates the NifA-3xFlag protein (~64 KDa), while the light grey thick arrow indicates the Fnr1-3xFlag protein (~34K Da). A representative result is show from three independent protein extract preparations.Back to article page