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Construction of R. leguminosarum GLF1 genomic nifA::lacZ fusion. A 6.1-kb SalI/XbaI fragment spanning the region from ΔnifH to nifA was cloned in suicide vector pVIK112 upstream of a lacZ reporter gene, generating plasmid pVIKM1. Single homologous recombination between the cloned fragment and genomic DNA resulted in integration of the entire pVIKM1 plasmid in the genome. The resulting GLF1 strain harbored a duplication of the cloned DNA fragment and a genomic nifA::lacZ fusion.
