Cloning, Expression and Location of the Gene for Phospho---galactosidase Free

Abstract

Summary: Genes for lactose catabolism and proteinase production in 712 are encoded by a 56.5 kb metabolic plasmid, pLP712. A lactose mini-plasmid of only 23.7 kb, pMG820, was constructed by introducing two deletions into pLP712, and was cloned as two segments of DNA into the vector pAT153 using restriction endonuclease I. The lactose genetic region of pLP712, which has been defined by deletion and restriction mapping, was cut into two parts by this process. When the smaller 10.8 kb segment of pMG820 DNA was present, the key lactic streptococcal lactose splitting enzyme, phospho---galactosidase, was expressed in The gene for phospho---galactosidase was more precisly located by introducing a series of deletions into cloned DNA by manipulations and then assaying for enzyme activity. The presence of this phospho---galactosidase activity was correlated with the production of a 58kDa S-labelled protein both by minicells and after coupled transcription and translation of cloned DNA. The product of a second gene, a 37 kDa protein (“protein X”), and a possible truncated phospho---galactosidase protein of 16 kDa were also detected in minicells.

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1986-02-01
2024-03-29
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References

  1. Bissett D. L., Anderson R. L. 1974; Lactose and D-galactose metabolism in group N streptococci: presence of enzymes for both the D-galactose-1-phosphate and D-tagatose-6-phosphate pathways. Journal of Bacteriology 117:318–320
    [Google Scholar]
  2. Bradford M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein, utilizing the principle of protein-dye binding. Analytical Biochemistry 12:248–254
    [Google Scholar]
  3. Calmes R. 1978; Involvement of phosphoenolpyru-vate in the catabolism of caries-conducive disaccharides by Streptococcus mutans: lactose transport. Infection and Immunity 19:934–942
    [Google Scholar]
  4. Collins J. 1979; Cell-free synthesis of proteins coding for mobilization functions of ColEl and transposition functions of Tn3. Gene 6:29–42
    [Google Scholar]
  5. Crow V. L, Davey G. P., Pearce L. E., Thomas T. D. 1983; Plasmid linkage of the D-tagatose-6-phosphate pathway in Streptococcus lactis: effect on lactose and galactose metabolism. Journal of Bacteriology 153:76–83
    [Google Scholar]
  6. DeVries J. K., Zubay G. 1967; DNA-directed peptide synthesis. II. The synthesis of the α fragment of the enzyme β-galactosidase. Proceedings of the National Academy of Sciences of the United States of America 57:1010–1012
    [Google Scholar]
  7. Dugaiczyk A., Boyer H. W. 1975; Ligation of EcoRI endonuclease-generated DNA fragments into linear and circular structures. Journal of Molecular Biology 96:171–184
    [Google Scholar]
  8. Gasson M. J. 1980; Production, regeneration and fusion of protoplasts in lactic streptococci. FEMS Microbiology Letters 9:99–102
    [Google Scholar]
  9. Gasson M. J. 1980; Plasmid complements of Streptococcus lactis NCD0 712 and other lactic streptococci after protoplast-induced curing. Journal of Bacteriology 154:1–9
    [Google Scholar]
  10. Hamilton I. R., Lo G. Y. C. 1978; Co-induction of β-galactosidase and the lactose-P-enoipyruvate phosphotransferase system in Streptococcus salivarius and Streptococcus mutans. . Journal of Bacteriology 136:900–908
    [Google Scholar]
  11. Heller K., Roschenthaler R. 1978; βd-phosphogalactosidase galactohydrolase of Streptococcus faecalis and the inhibition of its synthesis by glucose. Canadian Journal of Microbiology 24:512–519
    [Google Scholar]
  12. Hengstenberg W., Penberthy W. K, Hill K. L., Morse M. L. 1969; Phosphotransferase system of Staphylococcus aureus: its requirement for the accumulation and metabolism of galactosides. Journal of Bacteriology 99:383–388
    [Google Scholar]
  13. Humphreys G. O., Willshaw G. A., Anderson E. S. 1975; A simple method for the preparation of large quantities of pure plasmid DNA. Biochimica et biophysica acta 383:457–463
    [Google Scholar]
  14. Laemmli U. K., Favre M. 1973; Maturation of the head of bacteriophage T4.I. DNA packaging events. Journal of Molecular Biology 80:575–599
    [Google Scholar]
  15. Lee L. L., Hansen J. B., Jagusztyn-Krynicka E. K., Chassy B. M. 1982; Cloning and expression of the βd-phosphogalactoside galactohydrolase gene of Lactobacillus casei in Escherichia coli K12. Journal of Bacteriology 152:1138–1146
    [Google Scholar]
  16. McKay L. L., Walter L. A., Sandine W. E., Elliker P. R. 1969; Involvement of phosphoenol-pyruvate in lactose utilization by group N streptococci. Journal of Bacteriology 99:603–610
    [Google Scholar]
  17. McKay L. L., Walter L. A., Sandine W. E., Elliker P. R. 1970; Mechanisms of lactose utilization by lactic acid streptococci: enzymatic and genetic analyses. Journal of Bacteriology 102:804–809
    [Google Scholar]
  18. Okamoto T., Morichi T. 1979; Distribution of β-galactosidase and β-phosphogalactosidase activity among lactic streptococci. . Agricultural and Biological Chemistry 43:2389–2390
    [Google Scholar]
  19. Premi L., Sandine W. E., Elliker P. R. 1972; Lactose-hydrolysing enzymes of Lactobacillus species. . Applied Microbiology 24:51–57
    [Google Scholar]
  20. Radloff R., Bauer W., Vinograd J. 1967; A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proceedings of the National Academy of Sciences of the United States of America 57:1514–1521
    [Google Scholar]
  21. Reeve J. N. 1977; Bacteriophage infection of minicells. Molecular and General Genetics 158:73–79
    [Google Scholar]
  22. Reeve J. N. 1979; Use of minicells for bacteriophage-directed polypeptide synthesis. Methods in Enzymology 68:493–503
    [Google Scholar]
  23. Zubay G. 1973; In vitro synthesis of protein in microbial systems. Annual Review of Genetics 7:267–287
    [Google Scholar]
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