Summary
We have worked out conditions for the study of competence development and genetic transformation in Streptococcus oralis NCTC 11427 (type strain), a species that contains choline in the cell wall. The peak of competence was found at the early exponential phase of growth and the optimal conditions for transformation were achieved with shuttle plasmids prepared from S. pneumoniae or from Escherichia coli serving as donor DNA. Transformation with dye-bouyant density gradient purified plasmid preparations followed first-order kinetics. The pneumococcal amidase can be expressed in S. oralis harbouring a plasmid carrying the lytA gene. This enzyme lysed the cell wall of the transformed cell in the presence of detergents.
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References
Barany F, Tomasz A (1980) Genetic transformation of Streptococcus pneumoniae by heterologous plasmid deoxyrbonucleic acid. J Bacteriol 144:698–709
Barany F, Boeke JD, Tomasz A (1982) Staphylococcal plasmids that replicate and express erythromycin resistance in both Streptococcus pneumoniae and Escherichia coli. Proc Natl Acad Sci USA 79:2991–2995
Chen JD, Morrison DA (1987) Cloning of Streptococcus pneumoniae DNA fragments in Escherichia coli requires vectors protected by strong transcriptional terminators. Gene 55:179–187
Espinosa M, López P, Perez-Ureña MT, Lacks SA (1982) Interspecific plasmids transfer between S. pneumoniae and B. subtilis. Mol Gen Genet 188:195–201
García E, Ronda C, García JL, López R (1985a) A rapid procedure to detect the autolysin phenotype in Streptococcus pneumoniae. FEMS Microbiol Lett 29:77–81
García E, García JL, Ronda C, López R (1985b) Cloning and expression of the pneumococcal autolysin gene in Escherichia coli. Mol Gen Genet 201:225–230
García P, García JL, García E, López R (1986) Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli. Gene 43:265–272
García JL, García E, Arrarás A, García P, Ronda C, López R (1987) Cloning, purification, and biochemical properties of the pneumococcal bacteriophage Cp-1 lysin. J Virol 61:2573–2580
García E, García JL, García P, Arrarás A, Sánchez-Puelles JM, López R (1988a) Molecular evolution of lytic enzymes of Streptococcus pneumoniae and its bacteriophages. Proc Natl Acad Sci USA 85:914–918
García JL, García E, Sánchez-Puelles JM, López R (1988b) Identification of a lytic enzyme of Clostridium acetobutylicum that degrades choline-containing pneumococcal cell walls. FEMS Microbiol Lett 52:133–138
Höltje JV, Tomasz A (1976) Purification of the pneumococcal Nacetylmuramyl-L-alanine amidase to biochemical homogeneity. J Biol Chem 251:4199–4207
Kilpper-Bälz R, Wenzig P, Schleifer KH (1985) Molecular relationships and classification of some viridans streptococci as Streptococcus oralis and amended description of Streptococcus oralis (Bridge and Sneath 1982). Int J Syst Bacteriol 35:482–488
Lacks SA, López P, Greenberg B, Espinosa M (1986) Identification and analysis of genes for tetracycline resistance and replication functions in the broad-host-range plasmid pLS1. J Mol Biol 192:753–765
Lindler LE, Macrina FL (1986) Characterization of genetic transformation in Streptococcus mutants by using a novel high-efficiency plasmid marker rescue system. J Bacteriol 166:658–665
López R, Ronda C, García P, Escarmís C, García E (1984) Restriction cleavage maps of the DNAs of Streptococcus pneumoniae bacteriophages containing protein covalently bound to their 5′ ends. Mol Gen Genet 197:67–74
Macrina FL, Jones KR, Welch RA (1981) Transformation of Streptococcus sanguis with monomeric pVA736 plasmid deoxyribonucleic acid. J Bacteriol 146:826–830
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Mosser JL, Tomasz A (1970) Choline-containing teichoic acid as a structural component of pneumococcal cell wall and its role in sensitivity of lysis by an autolytic enzyme. J Biol Chem 245:287–298
Perry D, Wondrack LM, Kuramitsu HK (1983) Gentic transformation of putative cariogenic properties Streptococcus mutans. Infect Immun 41:722–727
Ronda C, García JL, García E, Sánchez-Puelles JM, López R (1987) Biological role of the pneumococcal amidase. Cloning of the lytA gene in Streptococcus pneumoniae. Eur J Biochem 164:621–624
Sánchez-Puelles JM, Ronda C, García JL, García P, López R, García E (1986) Searching for autolysin functions. Characterization of a pneumococcal mutant deleted in the lytA gene. Eur J Biochem 158:289–293
Saunders JR, Guild WR (1980) Properties and transformation activities of two plasmids in Streptococcus pneumoniae. Mol Gen Genet 180:573–587
Stassi DL, Lacks SA (1982) Effect of strong promoters on the cloning on Escherichia coli of DNA fragments from Streptococcus pneumoniae. Gene 18:319–328
Tomasz A (1967) Choline in the cell wall of a bacterium: Novel type of polymer-linked choline in pneomococcus. Science 157:694–697
Tomasz A (1970) Cellular metabolism in genetic transformation of pneumococci: Requirement for protein synthesis during induction of competence. J Bacteriol 101:860–871
Tomasz A, Hochtkiss RD (1964) Regulation of the transformability of pneomococcal cultures by macromolecular cell products. Proc Natl Acad Sci USA 51:480–487
Westergren G, Emilson CG (1983) Prevalence of transformable Streptococcus mutans in human dental plaque. Infect Immun 41:1386–1388
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Ronda, C., García, J.L. & López, R. Characterization of genetic transformation in Streptococcus oralis NCTC 11427: Expression of the pneumococcal amidase in S. oralis using a new shuttle vector. Mol Gen Genet 215, 53–57 (1988). https://doi.org/10.1007/BF00331302
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DOI: https://doi.org/10.1007/BF00331302