1887

Abstract

A rapid and efficient method of transforming by electroporation is described. A number of factors are important in determining the level of transformation obtained. Cells grown in the presence of glycine and isonicotinic acid hydrazide and harvested in early exponential growth phase were much easier to transform. The recovery medium on which transformants were isolated also had a significant effect on the number of transformants obtained because cells were osmotically or electrochemically sensitive following electroporation. Transformation efficiencies of up to 5 × 10 transformants per μg plasmid DNA with homologously derived DNA and 2 × 10 transformants per μg of DNA derived from were obtained.

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/content/journal/micro/10.1099/00221287-136-2-255
1990-02-01
2022-01-21
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References

  1. Allen S. P., Blaschek H. P. 1988; Electroporation-induced transformation of intact cells of Clostridium perfringens . Applied and Environmental Microbiology 54:2322–2324
    [Google Scholar]
  2. Best G. R., Britz M. L. 1986; Facilitated protoplasting in certain auxotrophic mutants of Corynebacterium glutamicum . Applied Microbiology and Biotechnology 23:288–293
    [Google Scholar]
  3. Britz M. L., Best G. R. 1986; Expression of chloramphenicol resistance specified by plasmid pHY416 hosted in Corynebacterium glutamicum . Current Microbiology 14:13–17
    [Google Scholar]
  4. Chassy B. M., Flickinger J. L. 1987; Transformation of Lactobacillus casei by electroporation. FEMS Microbiology Letters 44:173–177
    [Google Scholar]
  5. Dower W. J., Miller J. F., Ragsdale C. W. 1988; High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Research 16:6127–6145
    [Google Scholar]
  6. Hammes W., Schleifer K. H., Kandler O. 1973; Mode of action of glycine on the biosynthesis of peptidoglycan. Journal of Bacteriology 116:1029–1053
    [Google Scholar]
  7. Katsumata R., Ozaki A., Oka T., Furuya A. 1984; Protoplast transformation of glutamate-producing bacteria with plasmid DNA. Journal of Bacteriology 159:306–311
    [Google Scholar]
  8. Maekura R. 1984; Effect of isonicotinic acid hydrazide on mycolic acid synthesis in Nocardia rubra . Kekkaku 59:435–440
    [Google Scholar]
  9. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  10. Martin J. F., Santamaria R., Sandoval H., Delreal G., Mateos L. M., Gil J. A., Aguilar A. 1987; Cloning system in amino acid-producing corynebacteria. Biotechnology 5:137–147
    [Google Scholar]
  11. Miller J. H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. Miller J. F., Dower W. J., Tomkins L. S. 1988; High voltage electroporation of bacteria: genetic transformation of Campylobacter jejuni with plasmid DNA. Proceedings of the National Academy of Sciences of the United States of America 85:856–860
    [Google Scholar]
  13. Powell I. B., Achen M. G., Hillier A. J., Davidson B. E. 1988; A simple and rapid method for genetic transformation of lactic streptococci by electroporation. Applied and Environmental Microbiology 54:655–660
    [Google Scholar]
  14. Raleigh E. A. 1987; Restriction and modification in vivo by Escherichia coli K12. Methods in Enzymology 152:130–141
    [Google Scholar]
  15. Santamaria R., Gil J. A., Mesas J. M., Martin J. F. 1984; Characterization of an endogenous plasmid and development of cloning vectors and a transformation system in Brevibacterium lactofermentum . Journal of General Microbiology 130:2237–2246
    [Google Scholar]
  16. Santamaria R. I., Gil J. A., Martin J. F. 1985; High-frequency transformation of Brevibacterium lactofermentum by plasmid DNA. Journal of Bacteriology 162:463–467
    [Google Scholar]
  17. Serwold-Davis T. M., Groman N., Rabin M. 1987; Transformation of Corynebacterium diphtheriae, Corynebacterium ulcerans, Coryne- bacterium glutamicum, and Escherichia coli with the C. diphtheriae plasmid pN G2. Proceedings of the National Academy ofSciences of the United States of America 84:4964–4968
    [Google Scholar]
  18. Thierbach G., Schwarzer A., Puhler A. 1988; Transformation of spheroplasts and protoplasts of Corynebacterium glutamicum . Applied Microbiology and Biotechnology 29:356–362
    [Google Scholar]
  19. Tomiyasu I., Yano I. 1984; Isonicotinic acid hydrazide induced changes and inhibition in mycolic acid synthesis in Nocardia and related taxa. Archives of Microbiology 137:316–323
    [Google Scholar]
  20. Yeh P., Oreglia J., Prevots F., Sicard A. M. 1986; A shuttle vector system for Brevibacterium lactofermentum . Gene 47:301–306
    [Google Scholar]
  21. Yoshihama M., Higashiro K., Rao E. A., Akedo M., Shanabruch W. G., Follettie M. T., Walker G. C., Sinskey A. J. 1985; Cloning vector system for Corynebacterium glutamicum . Journal of Bacteriology 162:591–597
    [Google Scholar]
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