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. 1999 Jan 19;96(2):604–609. doi: 10.1073/pnas.96.2.604

Figure 3.

Figure 3

Expression of NtrB and derivatives. (A) Total proteins of cells harboring the indicated proteins were analyzed by using Western blot analysis. Lane 1, RB9132 (ΔntrB) carrying pTrc99B (vector); lane 2, YMC10 (NtrB+) grown in Ggln; lane 3, RB9132 carrying pgln62 (NtrB); lane 4, RB9132 carrying pB1 (NtrB); and lane 5, RB9132 carrying pBSHN1 (NtrBSHN). With the exception of YMC10, cells were grown under nitrogen-excess conditions (GLBgln). Expression of NtrBSHN was induced with 10 μM IPTG (lane 5); all other cells were grown in the absence of IPTG. (B) Western blot analysis of hybrid proteins containing the N-terminal domain of λ repressor. Cells (RB9132) were grown under nitrogen-excess conditions (GLBgln). Expression was induced with 10 μM IPTG except for λN-NtrBSH (50 μM IPTG). Lane 1, pJH391; lane 2, pFB1 (λN-NtrB); lane 3, pFBSHN1 (λN-NtrBSHN); lane 4, pFBSH1 (λN-NtrBSH); lane 5, pFBS1 (λN-NtrBS); lane 6, pFBHNG1 (λN-NtrBHNG); lane 7, pFBHN1 (λN-NtrBHN); and lane 8, pFBG1 (λN-NtrBG). (C) Overexpression of hybrid proteins containing the N-terminal domain of λ repressor. Crude cell extracts of RB9132 carrying the indicated plasmids were subjected to electrophoresis on an SDS/PAGE gel (12%) and visualized by staining with Coomassie blue. Expression was induced by the addition of 1 mM IPTG. Lane 1, molecular weight markers; lane 2, pJH391 (vector); lane 3, pFBHN1 (λN-NtrBHN); lane 4, pFBG1 (λN-NtrBG); lane 5, pFBSHN1 (λN-NtrBSHN); lane 6, pFBHNG1 (λN-NtrBHNG); lane 7, pFB1 (λN-NtrB); lane 8, pFBSH1 (λN-NtrBSH); and lane 9, pFBH1 (λN-NtrBH); and lane 10, pFBHNG287 (λN-NtrBHNG287).