1887

Abstract

Summary

Three previously reported Tn mutants of AM1, Cou-1, Cou-3 and Cou-6, which are able to grow on methylamine but not methanol, were characterized by biochemical analyses and complementation tests using two genomic libraries of AM1. We have designated the genes defective in these mutants as and and mapped the site of Tn insertion in each. Biochemical results showed that two of these methanol oxidation (Mox) mutants, Cou-1 and Cou-3, are phenotypically similar to the previously identified MoxE class of mutant while Cou-6 resembled the MoxD class. Complementation tests and mapping the site of the Tn insertions indicated that is another Mox gene linked to and that is linked to . The Tn insertion in mapped within the gene and therefore is the first detected mutation of the gene which was identified from expression studies. A MoxE mutant of AM1 was complemented by two different cosmid clones; one carried the gene and the other contained two gene clusters, and . Hybridization experiments indicated that the gene was not present on this latter clone, and it must therefore encode a gene capable of suppressing a MoxE mutation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-137-4-895
1991-04-01
2021-05-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/137/4/mic-137-4-895.html?itemId=/content/journal/micro/10.1099/00221287-137-4-895&mimeType=html&fmt=ahah

References

  1. Allen L. N., Hanson R. S. 1985; Construction of broad host range cloning vectors: identification of genes necessary for growth of Methylobacterium organophilum on methanol. Journal of Bacteriology 161:955–962
    [Google Scholar]
  2. Anderson D. J., Lidstrom M. E. 1988; The ‘mox FG’ region encodes four polypeptides in the methanol-oxidizing bacterium Methylobacterium sp. strain AM1. Journal of Bacteriology 170:2254–2262
    [Google Scholar]
  3. Anderson D. J., Morris C. J., Nunn D. N., Anthony C., Lidstrom M. E. 1990; Nucleotide sequences of the Methylobacterium extorquens sp. strain AM1 moxF and J genes involved in methanol oxidation. Gene 90:173–176
    [Google Scholar]
  4. Anthony C. 1986; The oxidation of methane and methanol by methylotrophs. Advances in Microbial Physiology 27:113–210
    [Google Scholar]
  5. Bastien C., Machlin S., Zhang Y., Donaldson K., Hanson R. S. 1989; Organization of genes required for the oxidation of methanol to formaldehyde in three type II methylotrophs. Applied and Environmental Microbiology 55:3124–3130
    [Google Scholar]
  6. Beardmore-Gray M., O’Keeffe D. T., Anthony C. 1983; The methanol: cytochrome c oxidoreductase activity of methylotrophs. Journal of General Microbiology 129:923–933
    [Google Scholar]
  7. Boyer H. W., Roulland-Dussoix D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli. Journal of Molecular Biology 41:459–472
    [Google Scholar]
  8. Cohen S. N., Chang A. C. Y., Hsu L. 1972; Non-chromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proceedings of the National Academy of Sciences of the United States of America 692110–2114
    [Google Scholar]
  9. Day D. J., Nunn D. N., Anthony C. 1990; Characterisation of a novel soluble c-type cytochrome in a moxD mutant of Methylobacterium extorquens AM1. Journal of General Microbiology 136:181–188
    [Google Scholar]
  10. Ditta G., Schmidhauser T., Yakobson E., Lu P., Liang X., Finlay D., Guiney D., Helinski D. 1985; Plasmids related to the broad host range vector, pRK290, useful for gene cloning and monitoring gene expression. Plasmid 13:149–153
    [Google Scholar]
  11. Dunstan P. M., Drabble W. T., Anthony C. 1972; Microbial metabolism of C-1 and C-2 compounds: the involvement of glycolate in the metabolism of ethanol and of acetate by Pseudomonas AM1. Biochemical Journal 128:99–106
    [Google Scholar]
  12. Figurski D. H., Helinski D. R. 1979; Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proceedings of the National Academy of Sciences of the United States of America 761648–1652
    [Google Scholar]
  13. Francis R. T., Becker R. R. 1984; Specific indication of hemoproteins in polyacrylamide gels using a double staining process. Analytical Biochemistry 136:509–514
    [Google Scholar]
  14. Fukumori Y., Yamanaka T. 1987; The methylamine oxidising system of Pseudomonas AM1 reconstituted with purified components. Journal of Biochemistry 101:441–445
    [Google Scholar]
  15. Fulton G. L., Nunn D. N., Lidstrom M. E. 1984; Molecular cloning of a malyl CoA lyase gene from Pseudomonas sp. strain AM1, a facultative methylotroph. Journal of Bacteriology 160:718–723
    [Google Scholar]
  16. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. Journal of Molecular Biology 166:557–580
    [Google Scholar]
  17. Kim Y. M., Lidstrom M. E. 1989; Plasmid analysis in pink facultative methylotrophic bacteria using a modified acetone-alkaline hydrolysis method. FEMS Microbiology Letters 60:125–130
    [Google Scholar]
  18. Knauf V. C., Nester E. W. 1982; Wide host range cloning vectors: cosmid clone bank of Agrobacterium Ti plasmids. Plasmid 8:45–54
    [Google Scholar]
  19. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  20. Lidstrom M., Stirling D. I. 1990; Methylotrophs: genetics and commercial applications. Annual Review of Microbiology 44:27–57
    [Google Scholar]
  21. Lyon B. R., Kearney P. P., Sinclair M. I., Holloway B. W. 1988; Comparative complementation mapping of Methylophilus spp. using cosmid gene libraries and prime plasmids. Journal of General Microbiology 134:123–132
    [Google Scholar]
  22. Machlin S. M., Tam P. E., Bastien C. A., Hanson R. S. 1988; Genetic and physical analyses of Methylobacterium organophilum XX genes encoding methanol oxidation. Journal of Bacteriology 170:141–148
    [Google Scholar]
  23. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning, a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. Moore A. T., Nayudu M., Holloway B. W. 1983; Genetic mapping in Methylophilus methylotrophus AS1. Journal of General Microbiology 129:785–799
    [Google Scholar]
  25. Nayudu M., Holloway B. W. 1981; Isolation and characterization of R-plasmid variants with enhanced chromosomal mobilisation ability in Escherichia coli K12. Plasmid 6:53–66
    [Google Scholar]
  26. Nunn D. N., Anthony C. 1989; The nucleotide sequence and deduced amino acid sequence of the cytochrome c L gene of Methylobacterium extorquens AM1, a novel class of c-type cyto-chrome. Biochemical Journal 256:673–676
    [Google Scholar]
  27. Nunn D. N., Lidstrom M. E. 1986a; Isolation and complementation analysis of 10 methanol oxidation mutant classes and identification of the methanol dehydrogenase structural gene of Methylobacterium sp. strain AM1. Journal of Bacteriology 166:581–590
    [Google Scholar]
  28. Nunn D. N., Lidstrom M. E. 1986b; Phenotypic characterisation of 10 methanol oxidation mutant classes in Methylobacterium sp. strain AM1. Journal of Bacteriology 166:591–597
    [Google Scholar]
  29. Nunn D. N., Day D. J., Anthony C. 1989; The second subunit of methanol dehydrogenase of Methylobacterium extorquens AM1. Biochemical Journal 260:875–862
    [Google Scholar]
  30. O’Keeffe D. T., Anthony C. 1980; The two cytochromes c in the facultative methylotroph Pseudomonas AM1. Biochemical Journal 192:411–419
    [Google Scholar]
  31. Peel D., & Quayle J. R. 1961; Microbial growth on C, compounds. I. Isolation and characterization of Pseudomonas AM1. Biochemical Journal 81:465–169
    [Google Scholar]
  32. Rigby P., Dickmann M., Rhodes C., Berg P. 1977; Labelling DNA to high specific activity in vitro by nick translation with DNA polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  33. Scott K. F., Rolfe B. G., Shine J. 1981; Biological nitrogen fixation: primary structure of the Klebsiella pneumoniae nifH and nifD genes. Journal of Molecular and Applied Genetics 1:71–81
    [Google Scholar]
  34. Simon R., Priefer U., Puhler A. 1983; A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in gram negative bacteria. Bio/Technology 1:784–791
    [Google Scholar]
  35. Smith G. E., Summers M. D. 1980; The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl-paper. Analytical Biochemistry 109:123–129
    [Google Scholar]
  36. Stone S., Goodwin P. M. 1989; Characterisation and complemen-tation of mutants of Methylobacterium AM1 which are defective in C-1 assimilation. Journal of General Microbiology 135:227–235
    [Google Scholar]
  37. Tatra P. K., Goodwin P. M. 1983; R-plasmid mediated chromosome mobilization in the facultative methylotroph Pseudomonas AM1. Journal of General Microbiology 129:2629–2632
    [Google Scholar]
  38. Tatra P. K., Goodwin P. M. 1985; Mapping of some genes involved in C-1 metabolism in the facultative methylotroph Methylobacterium sp. strain AM1 (Pseudomonas AM1). Archives of Microbiology 143:169–177
    [Google Scholar]
  39. Vogel H. J., BONNER D. M. 1956; Acetylornithase of Escherichia coli: partial purification and some properties. Journal of Biological Chemistry 218:97–106
    [Google Scholar]
  40. Whitta S., Sinclair M. I., Holloway B. W. 1985; Transposon mutagenesis in Methylobacterium AM1 (Pseudomonas AM1). Journal of General Microbiology 131:1547–1549
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-137-4-895
Loading
/content/journal/micro/10.1099/00221287-137-4-895
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error