1887

Abstract

We previously reported the cloning of a 1·6 kb III fragment (containing the junction of the repeating unit) from chromosomal DNA of strain B7 in which tandem amplification of a 16 kb region occurred, and the induction of B7-type gene amplification by competence transformation with this cloned fragment. Based on this result, we designed, on the chromosome, a gene amplification of the 22 kb repeating unit containing the a-amylase structural gene (), the tunicamycin-resistance gene () and the shikimate kinase structural gene (). We cloned only two short DNA fragments from both termini of the 22 kb region, constructed a junction structure of the designed repeating unit on pBR327 and transformed a wild-type strain by this constructed plasmid. As a result, we succeeded in obtaining tunicamycin-resistant (Tm) transformants in which the designed gene amplification of 22 kb occurred on the chromosome. The Tm transformants showed high productivity of -amylase and shikimate kinase. The copy number of the repeating unit was estimated to be 10–20. This system may provide an effective means of amplifying long (> 20 kb) DNA regions on the chromosome.

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1988-01-01
2021-08-02
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References

  1. Albertini A.M., Galizzi A. 1985; Amplification of a chromosomal region in Bacillus subtilis. Journal of Bacteriology 162:1203–1211
    [Google Scholar]
  2. Altenbuchner J., Cullum J. 1984; DNA amplification and an unstable arginine gene in Streptomyces lividans 66. Molecular and General Genetics 195:134–138
    [Google Scholar]
  3. Anagnostopoulos C., Spizizen J. 1961; Requirements for transformation in Bacillus subtilis. Journal of Bacteriology 81:741–746
    [Google Scholar]
  4. Anderson R.P., Roth J.R. 1977; Tandem genetic duplications in phage and bacteria. Annual Review of Microbiology 31:473–505
    [Google Scholar]
  5. Deretic V., Tomasek P., Darzins A., Chakrabarty A.M. 1986; Gene amplification induces mucoid phenotype in rec-2 Pseudomonas aeruginosa exposed to kanamycin. Journal of Bacteriology 165:510–516
    [Google Scholar]
  6. Edlund T., Normark S. 1981; Recombination between short DNA homologies causes tandem duplication. Nature; London: 292269–271
    [Google Scholar]
  7. Fishman S.E., Hershberger C.L. 1983; Amplified DNA in Streptomyces fradiae. Journal of Bacteriology 155:459–466
    [Google Scholar]
  8. Furusato T., Yamane K., Hashiguchi K., Tanimoto A., Harada S., Mori M., Yoda K., Yamasaki M., Tamura G. 1986; recE- dependent gene amplification induced by a tunicamycin resistant mutation (tmrA 7) in Bacillus subtilis. Agricultural and Biological Chemistry 50:1771–1775
    [Google Scholar]
  9. Fuwa H. 1954; A new method for microdetermination of amylase activity by the use of amylose as the substrate. Journal of Biochemistry 41:583–603
    [Google Scholar]
  10. Goldfarb D.S., Doi R.H., Rodriguez R.L. 1981; Expression of Tn9-derived chloramphenicol resistance in Bacillus subtilis. Nature; London: 24309–311
    [Google Scholar]
  11. Gutterson N.I., Koshland D.E.Jr 1983; Replacement and amplification of bacterial genes with sequences altered in vitro. Proceedings of the National Academy of Sciences of the United States of America 80:4894–4898
    [Google Scholar]
  12. Hashiguchi K., Tanimoto A., Nomura S., Yamane K., Yoda K., Harada S., Mori M., Furusato T., Takatsuki A., Yamasaki M., Tamura G. 1986; Amplification of the amyE-tmrB region on the chromosome in tunicamycin-resistant cells of Bacillus subtilis. Molecular and General Genetics 204:36–43
    [Google Scholar]
  13. Hass M., Yoshikawa H. 1969; Defective bacteriophage PBSH in Bacillus subtilis.2. Intracellular development of the induced prophage. Journal of Virology 3:248–260
    [Google Scholar]
  14. JanniÈre L., Niaudet B., Pierre E., Ehrlich S.D. 1985; Stable gene amplification in the chromosome of Bacillus subtilis. Gene 40:47–55
    [Google Scholar]
  15. Maniatis T., Fritsch E.F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  16. Mori M., Tanimoto A., Yoda K., Harada S., Koyama N., Hashiguchi K., Obinata M., Yamasaki M., Tamura G. 1986; Essential structure in the cloned transforming DNA that induces gene amplification of the Bacillus subtilis amyE-tmrB region. Journal of Bacteriology 166:787–794
    [Google Scholar]
  17. Nakatsukasa W.M., Nester E.W. 1972; Regulation of aromatic amino acid biosynthesis in Bacillus subtilis 168. Journal of Biological Chemistry 247:5972–5979
    [Google Scholar]
  18. Niaudet B., JanniÈre L., Ehrlich S.D. 1985; Integration of linear, heterologous DNA molecules into the Bacillus subtilis chromosome: mechanism and use in induction of predictable rearrangements. Journal of Bacteriology 163:111–120
    [Google Scholar]
  19. Nomura S., Yamane K., Sasaki T., Yamasaki M., Tamura G., Maruo B. 1978; Tunicamycin-resistant mutants and chromosomal locations of mutational sites in Bacillus subtilis. Journal of Bacteriology 136:818–821
    [Google Scholar]
  20. Ono H., Hintermann G., Crameri R., Wallis G., Hutter R. 1982; Reiterated DNA sequences in a mutant strain of Streptomyces glaucescens and cloning of the sequence in Escherichia coli. Molecular and General Genetics 186:106–110
    [Google Scholar]
  21. Sargent M.G., Bennett M.F. 1985; Amplification of a major membrane-bound DNA sequence of Bacillus subtilis. Journal of Bacteriology 161:589–595
    [Google Scholar]
  22. Sasaki T., Yamasaki M., Maruo B., Yoneda Y., Yamane K., Takatsuki A., Tamura G. 1976; Hyperproductivity of extracellular α-amylase by a tunicamycin resistant mutant of Bacillus subtilis. Biochemical and Biophysical Research Communications 70:125–131
    [Google Scholar]
  23. Southern E.M. 1975; Detection of specific sequences among DNA fragments separated by gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  24. Takatsuki A., Arima K., Tamura G. 1970; Tunicamycin, a new antibiotic I. Isolation and characterization of tunicamycin. Journal of Antibiotics 24:215–223
    [Google Scholar]
  25. Tanaka T., Sakaguchi K. 1978; Construction of a recombinant plasmid composed of B.subtilis leucine genes and a B.subtilis (natto) plasmid: its use as cloning vehicle in B.subtilis 168. Molecular and General Genetics 165:269–276
    [Google Scholar]
  26. Tlsty T.D., Albertini A.M., Miller J.H. 1984; Gene amplification in the lac region of E.coli. Cell 37:217–224
    [Google Scholar]
  27. Trowsdale J., Anagnostopoulos C. 1975; Evidence for the translocation of a chromosome segment in Bacillus subtilis strains carrying the trpE26 mutation. Journal of Bacteriology 122886–898
    [Google Scholar]
  28. Vasseghi H., Claverys J.P. 1983; Amplification of a chimeric plasmid carrying an erythromycin-resistance determinant introduced into the genome of Streptococcus pneumoniae. Gene 21:285–292
    [Google Scholar]
  29. Wilson C.R., Morgan A.E. 1985; Chromosomal-DNA amplification in Bacillus subtilis. Journal of Bacteriology 163:445–453
    [Google Scholar]
  30. Yamazaki H., Ohmura K., Nakayama A., Takeichi Y., Otozai K., Yamasaki M., Tamura G., Yamane K. 1983; α-Amylase genes (amyR2 and amyE+) from α-amylase-hyperproducing Bacillus subtilis strain: molecular cloning and nucleotide sequences. Journal of Bacteriology 156:327–337
    [Google Scholar]
  31. Young M. 1984; Gene amplification in Bacillus subtilis. Journal of General Microbiology 130:1613–1621
    [Google Scholar]
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