Summary
Previously we isolated a Rhizobium japonicum DNA fragment that was homologous to at least one of the nitrogenase structural genes of Klebsiella pneumoniae (Hennecke 1981). The expression of several subfragments thereof was now tested in minicells of Escherichia coli by cloning them into suitable vectors in such a way that they were transcribed from the strong promoters of antibiotic resistance genes. Recombinant plasmid-encoded polypeptides were then identified on polyacrylamide gels. In addition, Tn5 insertion mutations were used to determine (i) the direction of transcription, (ii) the transcriptional organization, and (iii) the exact termination sites of genes.
Two polypeptides were identified as the β and α subunits of the R. japonicum nitrogenase component 1 (Rj1). Their respective genes, nifD and nifK, are organized in one transcriptional unit, and are transcribed in the order nif \({DK}\). Expression of component 2 protein (Rj2) has not been achieved. It is, therefore, an open question as to whether its corresponding gene, nifH, is located adjacent to nifDK as in K. pneumoniae.
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Communicated by J. Schell
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Fuhrmann, M., Hennecke, H. Coding properties of cloned nitrogenase structural genes from Rhizobium japonicum . Molec Gen Genet 187, 419–425 (1982). https://doi.org/10.1007/BF00332622
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DOI: https://doi.org/10.1007/BF00332622