The nitrogen fixation of Lignobacter K17 is plasmid mediated. Nif plasmid was transferred from Lignobacter to other bacterial species and the transposon Tn9 was inserted into it. The molecular weight of this plasmid designated pUCS101, is of 19.8 Mdal. In this study we constructed in vitro a hybrid plasmid (pUCS110) by ligating HindIII digests of pUCS101 nif:: Tn9 and of RP4. Next it was proved that pUCS110 is able to complement the total deletion of the nif region in Klebsiella pneumoniae. The 50 Mdal plasmid pUCS110 was not maintained stably in Escherichia coli recA+ as in E. coli recA-. After being transferred to K. pneumoniae, pUCS110 showed a tendency to generate plasmids of various size from 2.8 to 78 Mdal. Bacteria harbouring plasmids of various size classes were more resistant to chloramphenicol than K. pneumoniae (pUCS110). Altered cleavage patterns were found in derivatives of pUCS110. The obtained results suggest that translocation of the transposon Tn9 can be responsible for the instability of pUCS110.
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