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Identification, cloning, and sequencing of DNA essential for encapsulation of Streptococcus pneumoniae

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Abstract

This paper reports the cloning and sequencing of a region of DNA from Streptococcus pneumoniae serotype 3 surrounding transposon Tn916, insertion of which was previously shown to result in lack of expression of the extracellular capsule. Sequence analysis revealed that the transposon inserted into a consensus insertion site 71 bp from the 5′ end of the cloned fragment. Within the clone, 3′ downstream regions from two different pneumococcal lytA genes were identified, as well as a putative 194 AA open reading frame (ORF1). Moreover, two copies of the repeat element BOX, oriented in opposite directions, were located immediately 3′ of orf1. Within the region bounded by the first pair of internal sequencing primers, analysis revealed that the fragment amplified by PCR was always of the same size. Moreover, Southern blotting showed that for all serotypes examined to date, homology exists with the cloned fragment. These results indicate that this region of the chromosome is highly conserved and, taken together with other independently derived data, suggest that interruptions or deletions within this DNA lead to unencapsulation.

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Literature Cited

  1. Arrecubieta C, Lopez R, Garcia E (1994) Molecular characterization of cap3A, a gene from the operon required for the synthesis of the capsule of Streptococcus pneumoniae type 3: sequencing of mutations responsible for the unencapsulated phenotype and localization of the capsular cluster on the pneumococcal chromosome. J Bacteriol 176:6375–6383

    Google Scholar 

  2. Caillaud F, Courvalin P (1987) Nucleotide sequence of the ends of the conjugative shuttle transposon Tn1545. Mol Gen Genet 209:110–115

    Google Scholar 

  3. Clewell DB, Helinski DR (1970) Properties of deoxyribonucleic acid-protein relaxation complex and strand specificity of the relaxation event. Biochemistry 9:4428–4440

    Google Scholar 

  4. Diaz E, Lopez R, Garcia JL (1992) Role of the major pneumococcal autolysin in the atypical response of a clinical isolate of Streptococcus pneumoniae. J Bacteriol 174:5508–5515

    Google Scholar 

  5. Dillard JP, Yother J (1991) Analysis of Streptococcus pneumoniae sequences cloned into Escherichia coli: effect of promoter strength and transcription terminators. J Bacteriol 173:5105–5109

    Google Scholar 

  6. Garcia E, Garcia P, Lopez R (1993) Cloning and sequencing of a gene involved in the synthesis of the capsular polysaccharide of Streptococcus pneumoniae type 3. Mol Gen Genet 239:188–195

    Google Scholar 

  7. Garcia P, Garcia JL, Garcia E, Lopez R (1986) Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli. Gene 43:265–272

    Google Scholar 

  8. Gasc A-M, Kauc L, Barraille P, Sicard M, Goodgal S (1991) Gene localization, size, and physical map of the chromosome of Streptococcus pneumoniae. J Bacteriol 173:7361–7367

    Google Scholar 

  9. Hardy KJ, Peterlin BM, Atchison RE, Stobo JE (1985) Regulation of expression of the human interferon gene. Proc Soc Natl Acad Sci USA 82:8173–8177

    Google Scholar 

  10. Martin B, Humbert O, Camara M, Guenzi E, Walker J, Mitchell T, Andrew P, Prudhomme M, Alloing G, Hakenbeck R, Morrison DA, Boulnois GJ, Claverys J-P (1992) A highly conserved repeated DNA element located in the chromosome of Streptococcus pneumoniae. Nucleic Acids Res 20:3479–3483

    Google Scholar 

  11. Murphy E (1989) Transposable elements in gram-positive bacteria. In: Berg DE, Howe MM (eds) Mobile DNA. Washington, D.C.: American Society for Microbiology, pp 269–288

    Google Scholar 

  12. Musher DM (1992) Infections caused by Streptococcus pneumoniae: clinical spectrum, pathogenesis, immunity and treatment. Clin Infect Dis 14:801–809

    Google Scholar 

  13. Rubens CE, Wessels MR, Heggen LM, Kasper DL (1987) Transposon mutagenesis of type III group B Streptococcus: correlation of capsule expression with virulence. Proc Soc Natl Acad Sci USA 84:7208–7212

    Google Scholar 

  14. Sambrook EF, Fritsch J, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press

    Google Scholar 

  15. Sanchez-Puelles JM, Ronda C, Garcia JL, Garcia P, Lopez R, Garcia E (1986) Searching for autolysin functions: Characterization of a pneumococcal mutant deleted in the lytA gene. Eur J Biochem 158:289–293

    Google Scholar 

  16. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  17. Watson DA, Musher DM (1990) Interruption of capsule expression in Streptococcus pneumoniae serotype 3 by insertion of transposon Tn916. Infect Immun 58:3135–3138

    Google Scholar 

  18. Watson DA, Musher DM, Jacobson JW, Verhoef J (1993) A brief history of the pneumococcus in biomedical research: a panoply of scientific discovery. Clin Infect Dis 17:913–924

    Google Scholar 

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Watson, D.A., Kapur, V., Musher, D.M. et al. Identification, cloning, and sequencing of DNA essential for encapsulation of Streptococcus pneumoniae . Current Microbiology 31, 251–259 (1995). https://doi.org/10.1007/BF00298383

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