1887

Abstract

promoter-containing clones were isolated from a genomic DNA library constructed in the transcriptional-translational fusion vector pYUB76. The promoter-containing DNA fragments were identified in the surrogate host by expression of the promoterless reporter gene of pYUB76. The expression signals exhibited a wide range of strengths, as indicated by their corresponding β-galactosidase activities. Eight clones were sequenced and characterized further. Predicted open reading frames and codon usage were identified by computer analysis. Database searching for related sequences using the BLAST method revealed no homologies. Transcriptional activity was measured by slot-blot hybridization with steady-state RNA isolated from clones. Primer extension analysis identified the transcription start sites within the cloned fragments. The promoter regions characterized in this study were used to establish a consensus promoter sequence for consensus hexanucleotide sequences of TGMCGT and CGGCCS centred approximately 35 and 10 bp upstream from the transcription startpoints do not correspond to the consensus hexanucleotides of promoters.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-3-921
1997-03-01
2021-10-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/3/mic-143-3-921.html?itemId=/content/journal/micro/10.1099/00221287-143-3-921&mimeType=html&fmt=ahah

References

  1. Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. 1990; Basic local alignment search tool.. J Mol Biol 215:403–410
    [Google Scholar]
  2. Andersson S.G.E., Sharp P.M. 1996; Codon usage in the Mycobacterium tuberculosis complex.. Microbiology 142:915–925
    [Google Scholar]
  3. Barletta R.G., Kim D.D., Snapper S.B., Bloom B.R., Jacobs W.R. 1992; Identification of expression signals of the mycobacteriophages Bxbl, LI and TM4 using the Escherichia-Mycobacterium shuttle plasmids pYUB75 and pYUB76 designed to create translational fusions to the lacZ gene.. J Gen Microbiol 138:23–30
    [Google Scholar]
  4. Bashyam M.D., Tyagi A.K. 1994; An efficient and high yielding method for isolation of RNA from mycobacteria.. BioTechniques 17:834–836
    [Google Scholar]
  5. Bashyam M.D., Kaushal D., DasGupta S.K., Tyagi A.K. 1996; A study of the mycobacterial transcriptional apparatus: identification of novel features in promoter elements.. J Bacteriol 178:4847–4853
    [Google Scholar]
  6. Clark-Curtiss J.E., Jacobs W.R., Doherty M.A., Ritchie L.R., Curtiss R. III 1985; Molecular analysis of DNA and construction of genomic libraries of M.leprae. . J Bacteriol 161:1093–1102
    [Google Scholar]
  7. Cupples C.G., Miller J.H. 1988; Effect of amino acid substitutions at the active site in Escherichia coli β-galactosidase.. Genetics 120:637–644
    [Google Scholar]
  8. Dale J.W., Patki A. 1990; Mycobacterial gene expression and regulation.. In Molecular Biology of the Mycobacteria pp. 173–198 Edited by McFadden J. J. London: Surrey University Press;
    [Google Scholar]
  9. DasGupta S.K., Bashyam M.D., Tyagi A.K. 1993; Cloning and assessment of mycobacterial promoters by using a plasmid shuttle vector.. J Bacteriol 175:5186–5192
    [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Res 12:387–395
    [Google Scholar]
  11. Douglas A.L., Hatch T.P. 1996; Mutagenesis of the P2 promoter of the major outer membrane protein gene of Chlamydia trachomatis. . J Bacteriol 178:5573–5578
    [Google Scholar]
  12. Ehretsmann C.P., Carpousis A.J., Krisch H.M. 1992; mRNA degradation in procaryotes.. FASEB (Fed Am Soc Exp Biol) J 6:3186–3192
    [Google Scholar]
  13. El-Zaatari F.A.K., Naser S.A., Engstrand L, Graham D.Y. 1994; Identification and characterization of Mycobacterium paratuberculosis recombinant proteins expressed in E. coli. . Curr Microbiol 29:177–184
    [Google Scholar]
  14. Estrada-G I.C.E., Lamb F.I., Colston M.J., Cox R.A. 1988; Partial nucleotide sequence of 16S ribosomal RNA isolated from armadillo-grown Mycobacterium leprae. . J Gen Microbiol 134:1449–1453
    [Google Scholar]
  15. Foley-Thomas E.M., Whipple D.L., Bermudez L.E., Barletta R.G. 1995; Phage infection, transfection, and transformation of Mycobacterium avium complex and Mycobacterium paratuberculosis. . Microbiology 141:1173–1181
    [Google Scholar]
  16. Gilot P., De Kesel M., Machtelinckx L., Coene M., Cocito C. 1993; Isolation and sequencing of the gene coding for an antigenic 34-kilodalton protein of Mycobacterium paratuberculosis. . J Bacteriol 175:4930–4935
    [Google Scholar]
  17. Green E.P., Tizard M.L.V., Moss M.T., Thompson J., Winter-bourne D.J., McFadden J.J., Hermon-Taylor J. 1989; Sequence and charateristics of IS900, an insertion element identified in a human Crohn’s disease isolate of Mycobacterium paratuberculosis. . Nucleic Acids Res 17:9063–9073
    [Google Scholar]
  18. Hance A.J., Grandchamp B., Levy-Frebault V., Lecossier D., Rauzier J., Bocart D., Gicquel B. 1989; Detection and identification of mycobacteria by amplification of mycobacterial DNA.. Mol Microbiol 3:843–849
    [Google Scholar]
  19. Hawley D.K., McClure W.R. 1983; Compilation and analysis of Escherichia coli promoter DNA sequences.. Nucleic Acids Res 11:2237–2255
    [Google Scholar]
  20. Honoré N., Bergh S., Chanteau S. 15 other authors 1993; Nucleotide sequence of the first cosmid from the Mycobacterium leprae genome project: structure and function of the Rif-Str regions.. Mol Microbiol 7:207–214
    [Google Scholar]
  21. Kremer L., Baulard A., Estaquier J., Content J., Capron A., Locht C. 1995; Analysis of the Mycobacterium tuberculosis 85A antigen promoter region.. J Bacteriol 177:642–653
    [Google Scholar]
  22. McFadden J.J., Butcher P.D., Chiodini R.J., Hermon-Taylor J. 1987; Determination of genome size and DNA homology between unclassified Mycobacterium species isolated from patients with Crohn’s disease and other mycobacteria.. J Gen Microbiol 133:211–214
    [Google Scholar]
  23. McLaughlin J.R., Murray C.L., Rabinowitz J.C. 1981; Unique features in the ribosome binding site sequence of the Grampositive Staphylococcus aureus β-lactamase gene.. J Biol Chem 256:11283–11291
    [Google Scholar]
  24. Malakooti J., Wang S.P., Ely B. 1995; A consensus promoter sequence for Caulobacter crescentus genes involved in biosynthetic and housekeeping function.. J Bacteriol 177:4372–4376
    [Google Scholar]
  25. Miller J.H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  26. Moran C.P. Jr Lang N., LeGrice S.F.J., Lee G., Stephens M., Sonenshein A.L., Pero J., Losick R. 1982; Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. . Mol Gen Genet 186:339–346
    [Google Scholar]
  27. Murray A., Winter N., Lagranderie M., Hill D.F., Rauzier J., Timm J., Leclerc C., Moriarty K.M., Gheorghiu M., Gicquel B. 1992; Expression of Escherichia coli β-galactosidase in Mycobacterium bovis BCG using an expression system isolated from Mycobacterium paratuberculosis which induced humoral and cellular immune responses.. Mol Microbiol 6:3331–3342
    [Google Scholar]
  28. Muto A., Osawa S. 1987; The guanine and cytosine content of genomic DNA and bacterial evolution.. Proc Natl Acad Sci USA 84166–169
    [Google Scholar]
  29. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Sela S., Clark-Curtiss J. 1991; Cloning and characterization of the Mycobacterium leprae ribosomal RNA promoter in Escherichia coli. . Gene 98:123–127
    [Google Scholar]
  31. Snapper S.B., Melton R.E., Mustafa S., Kieser T., Jacobs W.R. 1990; Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. . Mol Microbiol 4:1911–1919
    [Google Scholar]
  32. Stevenson K., Inglis N.F., Rae B., Donachie W., Sharp J.M. 1991; Complete nucleotide sequence of a gene encoding the 70 kd heat shock protein of Mycobacterium paratuberculosis. . Nucleic Acids Res 19:4552
    [Google Scholar]
  33. Stover C.K., de la Cruz V.F., Fuerst T.R. 11 other authors 1991; New use of BCG for recombinant vaccines.. Nature 351:456–460
    [Google Scholar]
  34. Stover K., Bansal G.P., Hanson M.S. 7 other authors 1993; Protective immunity elicited by recombinant bacille-Calmette- Guerin (BCG) expressing outer surface protein A (Osp A) lipoprotein: a candidate lyme disease vaccine.. J Exp Med 178:197–209
    [Google Scholar]
  35. Tartof K.D., Hobbs C.A. 1987; Improved media for growing plasmid and cosmid clones.. Bethesda Res Lab Focus 9:12
    [Google Scholar]
  36. Thole J.E.R., Dauwerse H.G., Das P.K., Groothjuis D.G., Schouls L.M., van Embden J.D.A. 1985; Cloning of Mycobacterium bovis BCG DNA and expression of antigens in Escherichia coli. . Infect Immun 50:800–806
    [Google Scholar]
  37. Thomas T.J., Thoen C.O., Andrews R.E. 1992; Molecular cloning and characterization of Mycobacterium paratuberculosis promoters in Escherichia coli. . Vet Microbiol 32:351–362
    [Google Scholar]
  38. Timm J.M., Perilli G., Uez C. 9 other authors 1994; Transcription and expression analysis using lacZ and phoA gene fusions of Mycobacterium fortuitum β-lactamase genes cloned from a natural isolate and a high level β-lactamase producer.. Mol Microbiol 12:491–504
    [Google Scholar]
  39. Tinoco I., Borer P.N., Dengler B., Levine M.D., Uhlenbeck O.C., Crothers D.M., Gralla J. 1973; Improved estimation of secondary structure in ribonucleic acids.. Nature New Biol 246:40–41
    [Google Scholar]
  40. Whipple D.L., Le Febvre R.B., Andrews R.E. Jr Thiermann A.B. 1987; Isolation and analysis of restriction endonuclease digestive patterns of chromosomal DNA from Mycobacterium paratuberculosis and other Mycobacterium species.. J Clin Microbiol 25:1511–1515
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-3-921
Loading
/content/journal/micro/10.1099/00221287-143-3-921
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