Standard cloning procedures were used to produce a strain constitutively expressing NifAl and the following constructs on suicide plasmids: amtB::¥LmR, amtYwGmR, glnK::GmR. Plasmids were introduced into R. capsulatus using conjugation and mutant strains selected and verified. Nitrogenase activity was measured by the acetylene reduction method and ADP-ribosylation of Fe-protein was detected using Western blots as previously described (Yakunin et al. 1999).
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