Transposon Tnmob, a Tn derivative containing the mobilization site of plasmid RP4, was introduced into strain 128C53 on the suicide vector pSup5011. Transposon insertions into the plasmids of strain 128C53 were selected after mating the mutagenized population with a non-nodulating recipient. In this way, a tenfold enrichment was effected for plasmid-linked mutations. Transconjugants that had received mutagenized symbiotic plasmids were applied individually to the roots of pea seedlings and, after three weeks, the resulting nodules were screened for the absence of hydrogenase activity. Eight hydrogenase-deficient (Hup) mutants of strain 128C53 were isolated. Physical and genetic analyses suggest that each Hup mutant was due to a single insertion of Tn5-mob into the symbiotic plasmid, pRL6JI. All mutations were suppressible by pHUl, a cosmid clone carrying some of the genes of .


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