Protoplast Transformation of Glutamate-producing Bacteria with Plasmid DNA

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1984 Jul 1

PMID 6145700

Citations 34

Authors

R Katsumata

A Ozaki

T Oka

A Furuya

Affiliations

Soon will be listed here.

Abstract

A method for polyethylene glycol-induced protoplast transformation of glutamate-producing bacteria with plasmid DNA was established. Protoplasts were prepared from cells grown in the presence of penicillin by treatment with lysozyme in a hypertonic medium. The concentration of penicillin during growth affected the efficiency of formation, regeneration, and polyethylene glycol-induced DNA uptake of protoplasts. Regeneration of protoplasts was accomplished on a hypertonic agar medium containing sodium succinate and yeast extract. The spectinomycin and streptomycin resistance plasmid pCG4, originally from Corynebacterium glutamicum T250, could transform various glutamate-producing bacteria such as C. glutamicum, Corynebacterium herculis, Brevibacterium flavum, and Microbacterium ammoniaphilum. The plasmid was structurally unchanged and stably maintained in new hosts. The transformation frequency of most competent protoplasts with pCG4 DNA isolated from primary transformants was high (ca. 10(6) transformants per microgram of covalently closed circular DNA) but was still two orders of magnitude below the frequency of transfection with modified DNA of the bacteriophage phi CGI. The difference was ascribed to the involvement of regeneration in transformation.

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References

Radloff R, BAUER W, Vinograd J . A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells. Proc Natl Acad Sci U S A. 1967; 57(5):1514-21. PMC: 224502. DOI: 10.1073/pnas.57.5.1514. View

Kawabe H, Mitsuhashi S . Inactivation of dihydrostreptomycin by Staphylococcus aureus. Jpn J Microbiol. 1971; 15(6):545-8. DOI: 10.1111/j.1348-0421.1971.tb00618.x. View

Guerry P, Leblanc D, Falkow S . General method for the isolation of plasmid deoxyribonucleic acid. J Bacteriol. 1973; 116(2):1064-6. PMC: 285495. DOI: 10.1128/jb.116.2.1064-1066.1973. View

Okanishi M, Suzuki K, Umezawa H . Formation and reversion of Streptomycete protoplasts: cultural condition and morphological study. J Gen Microbiol. 1974; 80(2):389-400. DOI: 10.1099/00221287-80-2-389. View

Humphreys G, Willshaw G, Anderson E . A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta. 1975; 383(4):457-63. DOI: 10.1016/0005-2787(75)90318-4. View

Landman O . Inhibitory protein controls the reversion of protoplasts and L forms of Bacillus subtilis to the walled state. J Bacteriol. 1977; 129(2):678-89. PMC: 234997. DOI: 10.1128/jb.129.2.678-689.1977. View

Hinnen A, Hicks J, Fink G . Transformation of yeast. Proc Natl Acad Sci U S A. 1978; 75(4):1929-33. PMC: 392455. DOI: 10.1073/pnas.75.4.1929. View

Bibb M, Ward J, Hopwood D . Transformation of plasmid DNA into Streptomyces at high frequency. Nature. 1978; 274(5669):398-400. DOI: 10.1038/274398a0. View

Baltz R . Genetic recombination in Streptomyces fradiae by protoplast fusion and cell regeneration. J Gen Microbiol. 1978; 107(1):93-102. DOI: 10.1099/00221287-107-1-93. View

10.

Chang S, Cohen S . High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979; 168(1):111-5. DOI: 10.1007/BF00267940. View

11.

Gabor M, HOTCHKISS R . Parameters governing bacterial regeneration and genetic recombination after fusion of Bacillus subtilis protoplasts. J Bacteriol. 1979; 137(3):1346-53. PMC: 218319. DOI: 10.1128/jb.137.3.1346-1353.1979. View

12.

Brown B, Carlton B . Plasmid-mediated transformation in Bacillus megaterium. J Bacteriol. 1980; 142(2):508-12. PMC: 294013. DOI: 10.1128/jb.142.2.508-512.1980. View

13.

Thompson C, Ward J, Hopwood D . DNA cloning in Streptomyces: resistance genes from antibiotic-producing species. Nature. 1980; 286(5772):525-7. DOI: 10.1038/286525a0. View

14.

Schiller J, GROMAN N, Coyle M . Plasmids in Corynebacterium diphtheriae and diphtheroids mediating erythromycin resistance. Antimicrob Agents Chemother. 1980; 18(5):814-21. PMC: 284096. DOI: 10.1128/AAC.18.5.814. View

15.

Martin P, Lohr J, Dean D . Transformation of Bacillus thuringiensis protoplasts by plasmid deoxyribonucleic acid. J Bacteriol. 1981; 145(2):980-3. PMC: 217207. DOI: 10.1128/jb.145.2.980-983.1981. View

16.

Imanaka T, Fujii M, Aramori I, Aiba S . Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis. J Bacteriol. 1982; 149(3):824-30. PMC: 216468. DOI: 10.1128/jb.149.3.824-830.1982. View

17.

Thompson C, Ward J, Hopwood D . Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol. 1982; 151(2):668-77. PMC: 220307. DOI: 10.1128/jb.151.2.668-677.1982. View

18.

Waxman D, Strominger J . Penicillin-binding proteins and the mechanism of action of beta-lactam antibiotics. Annu Rev Biochem. 1983; 52:825-69. DOI: 10.1146/annurev.bi.52.070183.004141. View