1887

Abstract

Summary: Expression studies on four members of the pMGA multigene family in A large family of related genes known as pMGA exists in the avian pathogen but only a single member of this family was previously found to be expressed in one strain of this bacterium. In this work two unrelated strains of were also shown by amino-terminal sequencing to express a unique pMGA polypeptide in both cases. To investigate pMGA gene selection in , mRNA expression was analysed in strain S6 using reverse transcription-PCR (RT-PCR and Northern blot techniques with probes for several members of the pMGA multigene family. It was shown that the pMGA message is 2·2 kb in size and is monocistronic. RT-PCR detected four different pMGA mRNA molecules but the relative yields were significantly affected by magnesium concentration. By quantitative Northern analysis, the relative abundances of the four pMGA mRNAs in S6 total RNA was determined: the pMGA1.1 mRNA predominated [1·88 ng (μg total RNA)] but at least three other pMGA genes were found to be transcribed but at much lower levels (20 to 40-fold lower). The pMGA1.1 mRNA is expressed at a level five times higher than the gene known to be one of the most abundantly expressed proteins in the prokaryoti cell. The start point of transcription for pMGA1.1 was determined and probable promoter assigned. From these data it appears likely that transcriptional control plays a major role in the selection of pMGA gene expression in the cell. S6

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-11-3005
1995-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/11/mic-141-11-3005.html?itemId=/content/journal/micro/10.1099/13500872-141-11-3005&mimeType=html&fmt=ahah

References

  1. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.. Anal Biocbem 162:156–159
    [Google Scholar]
  2. Church G.M., Gilbert W. 1984; Genomic sequencing.. Proc Natl Acad Sci USA 811991–1995
    [Google Scholar]
  3. Del Sal G., Manfidetti G., Schneider C. 1988; A one-tube plasmid DNA mini-preparation suitable for sequencing.. Nucleic Acids Res 16:9878
    [Google Scholar]
  4. Frey M.L, Anderson D.P. 1968; A medium for the isolation of avian mycoplasmas.. Am J Vet Res 29:2163–2171
    [Google Scholar]
  5. Hawley D.K., McClure W.R. 1983; Compilation and analysis of Escherichia coli promoter DNA sequences.. Nucleic Acids Res 11:2237–2255
    [Google Scholar]
  6. Higgins D.G., Bleasby A.J., Fuchs R. 1992; clustal v: improved software for multiple sequence alignment.. CABIOS 8:189–191
    [Google Scholar]
  7. Higgins P.A., Whithear K.G. 1986; Detection and differentiation of Mycoplasma gallisepticum and Mycoplasma synoviae antibodies in chicken serum using ELISA.. Avian Dis 30:160–168
    [Google Scholar]
  8. Jordan F.T.W. 1979; Avian mycoplasmas.. In The Mycoplasmas pp. 1–49 Tully J. G., Whitcomb R. F. Edited by New York:: Academic Press.;
    [Google Scholar]
  9. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  10. Loechel S., Inamine J.M., Hu P. 1989; Nucleotide sequence of the tuf gene from Mycoplasma gallisepticum. . Nucleic Acids Res 1710126
    [Google Scholar]
  11. Markham P.F., Glew M.D., Brandon M.R., Walker I.D., Whithear K.G. 1992; Characterization of a Major Hemagglutinin Protein from Mycoplasma gallisepticum. . Infect Immun 60:3885–3889
    [Google Scholar]
  12. Markham P.F., Glew M.D., Whithear K.G., Walker I.D. 1993; Molecular cloning of a member of the gene family that encodes pMGA, a hemagglutinin of Mycoplasma gallisepticum. . Infect Immun 61:903–909
    [Google Scholar]
  13. Markham P.F., Glew M.D., Sykes J.E., Bowden T.R., Pollocks T.D., Browning G.F., Whithear K.G., Walker I.D. 1994; The organisation of the multigene family which encodes the major cell surface protein, pMGA, of Mycoplasma gallisepticum. . FEBS Lett 325:347–352
    [Google Scholar]
  14. Matsudaira P. 1987; Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes.. J Biol Chem 262:10035–10038
    [Google Scholar]
  15. Muto A., Andochi Y., Yamao F., Tanaka R., Osawa S. 1992; Transcription and translation.. In Mycoplasmas: Molecular Biology and Pathogenesis pp. 331–347 Maniloff J., McElhaney R. N., Finch L. R., Baseman J. B. Edited by Washington, DC:: American Society for Microbiology.;
    [Google Scholar]
  16. Pedersen S., Bloch P.L., Reeh S., Neidhardt F.C. 1978; Patterns of protein synthesis in E. coli: a catalog of the amount of 140 individual proteins at different growth rates.. Cell 14:179–190
    [Google Scholar]
  17. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: A Eaboratory Manual, 2nd. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  18. Simecka J.W., Davis J.K., Davidson M.K., Ross S.E., Stadt-lander C.T.K.-H., Cassell G.H. 1992; Mycoplasma diseases of animals.. In Mycoplasmas: Molecular Biology and Pathogenesis pp. 391–415 Maniloff J., McElhaney R. N., Finch L. R., Baseman J. B. Edited by Washington, D.C:: ASM.;
    [Google Scholar]
  19. Sylvers L.A., Beresten S. 1993; Loss of resolution in gel electrophoresis of RNA: a problem associated with the presence of formaldehyde gradients.. Biotech 14:380–381
    [Google Scholar]
  20. Van De Putte P., Goosen N. 1992; DNA inversions in phages and bacteria.. Trends Genet 8:457–462
    [Google Scholar]
  21. van der Woude M.W., Braaten B.A., Low D.A. 1992; Evidence for global regulatory control of pilus expression in Escherichia coli by Lrp and DNA methylation: model building based on analysis of pap. . Mol Microbiol 6:2429–2435
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-11-3005
Loading
/content/journal/micro/10.1099/13500872-141-11-3005
Loading

Data & Media loading...

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