Summary
DNA from Clostridium acetobutylicum ABKn8 was partially digested with Sau3A and the fragments obtained were inserted into the unique BamHI site of the cloning vector pHV33. The recombinant plasmids were used to transform Escherichia coli HB101 with selection for ampicillin resistance. A collection of ampicillin-resistant, tetracycline-sensitive clones representative of the Clostridium acetobutylicum genome was made. The clones were shown to carry recombinant plasmids each containing an insert of 2 to 16 kb in size. Several of them complemented the HB101 proA2 or leuB6 auxotrophic mutations. The cloned sequences were shown by Southern blot hybridization to be homologous to the corresponding ABKn8 DNA fragments.
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References
Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–445
Anagnostopoulos C, Spizizen J (1961) Requirements for transformation in Bacillus subtilis. J Bacteriol 81:741–746
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472
Cameron JR, Panasenko SM, Lehman IR, Davis RW (1975) In vitro construction of bacteriophage lambda carrying segments of the Escherichia coli chromosome: selection of hybrids containing the gene for DNA ligase. Proc Natl. Acad Sci USA 72:3416–3420
Clarke L, Carbon J (1976) A colony bank containing synthetic Col El hybrid plasmids representative of the entire Escherichia coli genome. Cell 9:91–99
Dagert M, Ehrlich SD (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6:23–28
Efstathiou I, Truffaut N (1984) Clonage et expression de gènes de Clostridium acetobutylicum dans Escherichia coli. La génétique des micro-organismes industriels. Colloque Soc. Fse de Microbiologie, H Heslot (ed), SFM, pp 343–347
Ehrlich SD, Bursztyn-Pettegrew H, Stroynowski I, Lederberg J (1976) Expression of the thymidylate synthetase gene of the Bacillus subtilis bacteriophage ϕ-3-I in Escherichia coli. Proc Natl Acad Sci USA 73:4145–4149
Ish-Horowicz O, Burke JF (1981) Rapid and efficient cosmid cloning. Nucleic Acids Res 9:2989–2998
Ishii K, Kudo T, Honda H, Horikoshi K (1983) Molecular cloning of β-isopropylmalate dehydrogenase gene for Clostridium butryricum M588. Agric Biol Chem 47:2313–2317
Karube I, Urano N, Yamada T, Hirochika H, Sakaguchi K (1983) Cloning and expression of the hydrogenase gene from Clostridium butyricum in Escherichia coli. FEBS Lett 158:119–122
Mandel M, Higa A (1970) Calcium dependent bacteriophage DNA infection. J Mol Biol 53:159–162
Maniatis T, Hardison RC, Lacy E, Lauer J, O'Connel C, Quon D, Sim DK, Efstratiadis A (1978) The isolation of structural genes from libraries of eucaryotic DNA. Cell 15:687–701
Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218
Meyers JA, Sanchez D, Elwell LP, Falkow S (1976) Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J Bacteriol 127:1529–1537
Oultram JD, Young M (1985) Conjugal transfer of plasmid pAMB1 from Streptococcus lactis and Bacillus subtilis to Clostridium acetobutylicum. FEMS Microbiol Lett 27:129–134
Popoff MR, Truffaut N (1985) Survey of plasmids in Clostridium butyricum and Clostridium beijerinckii strains from different origins and different phenotypes. Curr Microbiol 12:151–156
Primrose SB, Ehrlich SD (1981) Isolation of plasmid deletion mutants and study of their instability Plasmid 6:193–201
Reysset G, Sebald M (1985) Conjugal transfer of plasmid-mediated antibiotic resistance from Streptococei to Clostridium acetobutylicum. Ann Microbiol (Inst Pasteur) 136B:275–282
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Sebald M, Costilow PN (1975) Minimal grwoth requirements for Clostridium perfringens and isolation of auxotrophic mutants. Appl Microbiol 29:1–6
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Tronik D (1982) Etudes physiologiques et génétiques d'une bactérie cellulolytique anaérobie thermophile: Clostridium thermocellum. Thesis 1982, Université Paris VII
Truffaut N, Sebald M (1983) Plasmid detection and isolation in strains of Clostridium acetobutylicum and related species. Mol Gen Genet 189:178–180
Ward JB, Zahler SA (1973) Genetic studies of leucine biosynthesis in Bacillus subtilis. J Bacteriol 116:719–726
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Efstathiou, I., Truffaut, N. Cloning of Clostridium acetobutylicum genes and their expression in Escherichia coli and Bacillus subtilis . Molec. Gen. Genet. 204, 317–321 (1986). https://doi.org/10.1007/BF00425516
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DOI: https://doi.org/10.1007/BF00425516