1887

Abstract

The fumarase () gene of 168 has been identified in a collection of λ phages carrying RI-generated fragments of DNA. Regions of the cloned DNA have been subcloned into plasmid vectors, and the ability of prophages and multicopy plasmids to complement and fumarase mutations has been examined. Two RI fragments of 1·5 and 5·1 kb are both required for fumarase expression in and The level of fumarase activity from a single copy of the gene expressed in is approximately one-tenth of that from the normal gene; the level is increased by expression from a pBR322-derived multicopy plasmid. The gene has been located within the cloned DNA by transposon mutagenesis and by expression studies, which have also identified a polypeptide of 49000 as the product of the gene. The properties of a truncated derivative of this polypeptide have indicated the direction of transcription of the gene.

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1984-11-01
2024-05-03
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References

  1. Anagnostopoulos C., Spizizen J. 1961; Requirements for transformation in Bacillus subtilis. Journal of Bacteriology 81:741–746
    [Google Scholar]
  2. Beeckmans S.A., Kanarek L. 1977; A new purification procedure for fumarase based on affinity chromatography. European Journal of Biochemistry 78:437–444
    [Google Scholar]
  3. Birnboim H.C.A., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1313–1524
    [Google Scholar]
  4. Carls R.A.A., Hanson R.S. 1971; Isolation and characterisation of tricarboxylic acid cycle mutants of Bacillus subtilis. Journal of Bacteriology 106:848–855
    [Google Scholar]
  5. De Lencastre H., Chak K.F., Piggot P.J. 1983; Use of the Escherichia coli transposon Tn 1000 (yd) to generate mutations in Bacillus subtilis DNA. Journal of General Microbiology 129:3203–3210
    [Google Scholar]
  6. Efstratiadis A., Vournakis J.N., Donis-Keller H., Chaconas G., Dougall D.K., Kafatos F.C. 1977; End labelling of enzymatically decapped mRNA. Nucleic Acids Research 4:4165–4174
    [Google Scholar]
  7. Guest J.R. 1981; Partial replacement of succinate dehydrogenase function by phage- and plasmid- specified fumaratereductase in Escherichia coli. Journal of General Microbiology 122:171–179
    [Google Scholar]
  8. Guest J.R., Roberts R.E. 1983; Cloning, mapping and expression of the fumarase gene of Escherichia coli K12. Journal of Bacteriology 153:588–596
    [Google Scholar]
  9. Guest J.R., Stephens P.E. 1980; Molecular cloning of the pyruvate dehydrogenase complex genes of Escherichia coliKI2. Journal of General Microbiology 121:277–292
    [Google Scholar]
  10. Guest J.R., Cole S.T., Jeyaseelan K. 1981; Organization and expression of the pyruvate dehydrogenase complex of Escherichia coli. Journal of General Microbiology 127:65–79
    [Google Scholar]
  11. Guyer M.S. 1978; The δε sequence of F is an insertion sequence. Journal of Molecular Biology 126:347–365
    [Google Scholar]
  12. Henner D.J., Hoch J.A. 1980; The Bacillus subtilis chromosome. Microbiological Reviews 44:57–82
    [Google Scholar]
  13. Hill R.L., Bradshaw R.A. 1969; Fumarase. Methods in Enzymology 13:91–99
    [Google Scholar]
  14. Lederberg E.M., Cohen S.N. 1974; Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. Journal of Bacteriology 119:1072–1074
    [Google Scholar]
  15. Miles J.S., Guest J.R. 1984; Complete nucleotide sequence of the fumarase gene, fumA, of Escherichia coli. Nucleic Acids Research 12:3631–3642
    [Google Scholar]
  16. Moir A. 1983; The isolation of λ transducing phages carrying the citG and gerA genes of Bacillus sttbtilis. Journal of General Microbiology 129:303–310
    [Google Scholar]
  17. Moir A., Smith D.A. 1983; The genetics of spore germination in Bacillus subtilis. In Fundamental and Applied Aspects of Spores pp. 89–100 Edited by Dring G.J., Ellar D.J., Gould G.W. London: Academic Press;
    [Google Scholar]
  18. Murray N.E., Brammar W.J., Murray K. 1977; Lambdoid phages (hat simplify the recovery of in vitro recombinants. Molecular and General Genetics 150:53–61
    [Google Scholar]
  19. Niaudet B., Ehrlich S.D. 1979; In vitro genetic labelling of Bacillus subtilis cryptic plasmid pH V400. Plasmid 2:48–58
    [Google Scholar]
  20. Primrose S.B., Ehrlich S.D. 1981; Isolation of plasmid deletion mutants and study of their instability. Plasmid 6:193–201
    [Google Scholar]
  21. Rutberg B., Hoch J.A. 1970; Citric acid cycle: gene-enzyme relationships in Bacillus subtilis. Journal of Bacteriology 104:826–833
    [Google Scholar]
  22. Sammons R.L., Anagnostopoulos C. 1982; Identification of a cloned DNA segment at a junction of chromosome regions involved in rearrangements in the trpE26 strains of Bacillus subtilis. FEMS Microbiology Letters 15:265–268
    [Google Scholar]
  23. Sancar A.A., Hak A.M., Rupp W.D. 1979; Simple method of identification of plasmid-coded proteins. Journal of Bacteriology 137:692–693
    [Google Scholar]
  24. Spencer M.E., Guest J.R. 1982; Molecular cloning of four tricarboxylic acid cycle genes of Escherichia coli. Journal of Bacteriology 151:542–552
    [Google Scholar]
  25. Warburg R.J., Moir A. 1981; Properties of a mutant of Bacillus subtilis 168 in which spore germination is blocked at a late stage. Journal of General Microbiology 124:243–253
    [Google Scholar]
  26. Weitzman P.J.D. 1981; Unity and diversity in some bacterial citric acid-cycle enzymes. Advances in Microbial Physiology 22:185–244
    [Google Scholar]
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