Construction of Clostridium butyricum hybrid plasmids and transfer to Bacillus subtilis

Luczak, Hania; Schwarzmoser, Hermine; Staudenbauer, Walter L.

doi:10.1007/bf00249941

Cited by 5 publications

(7 citation statements)

References 28 publications

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“…It has been classified by Gruss & Ehrlich (1989) in the pC194-type sub-group. Nevertheless, transformation of B. subtilis by pCTC511 strain BD224 occurred with poor efficiency as already observed by Collins et al (1985) and Luczak et al (1985), and plasmid DNA was barely detectable in isolates (see Fig. 6).…”

Section: Studies On the Segregational Stability Of The Two Strains Ofsupporting

confidence: 51%

“…In constrast, plasmid pCTC5 1 1, based on the clostridial pCBlOl replicon, was easily established in C. acetobutylicurn but not in B. subtilis (Collins et al, 1985;Luczak et al, 1985), and showed no segregational instability even in the parent strain NI-4081. This plasmid was shown to belong to the ssDNA family, but its replication strategy seems to be quite different from the Gram-positive plasmids of aerobic bacteria (Minton et al, 1990a, b).…”

Section: Discussionmentioning

confidence: 99%

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Stable inheritance of shuttle vectors based on plasmid pIM13 in a mutant strain of Clostridium acetobutylicum

Azeddoug

Hubert²,

Reysset³

1992

Journal of General Microbiology

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New shuttle vectors for Clostridium acefobufylicum were constructed, using as replicons the Gram-positive plasmid pIM13, and derivatives of the Gram-negative plasmid pBR322, including pUC19. These vectors transformed C. acefobufylicum at a high frequency (up to lo6 transformants per pg DNA) by PEG-mediated protoplast transformation. A mutant host strain, NI-4082, was isolated on the basis of its ability to maintain plasmid pIM13 stably in the absence of selection pressure. The shuttle vectors showed no segregational or structural instability in this mutant strain. Moreover, the results suggested a relationship between segregational instability and the multimerization of pIM13 in C. acefobufylicum The host/vector system described possessed all the properties required for efficient gene cloning in this species.

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Section: Studies On the Segregational Stability Of The Two Strains Ofsupporting

confidence: 51%

Section: Discussionmentioning

confidence: 99%

Stable inheritance of shuttle vectors based on plasmid pIM13 in a mutant strain of Clostridium acetobutylicum

Azeddoug

Hubert²,

Reysset³

1992

Journal of General Microbiology

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“…The method used to identify the replicons of pCB101 and pIP404 has not proved applicable to all clostridial plasmids. Restriction fragments derived from the C. butyricum plasmids pCB102 and pCB103 [111,65], and the C. paraputrificum plasraids pCP1 and pCP2 (T.J. Swinfield and N.P. Minton, unpublished) did not enable Rep-plasraids to replicate in B. subtilis.…”

Section: Indigenous Clostridial Plasmidsmentioning

confidence: 99%

“…Minton, unpublished) did not enable Rep-plasraids to replicate in B. subtilis. However, a pCP1derived plasmid was established in C. acetobutyli- [65,111]; d These vectors contain the oriT region of plasmid RK2; e pAT187 has been transferred to a wide spectrum of Gram-positive bacteria [74]; r pIM13 is a B. subtilis plasmid.…”

Section: Indigenous Clostridial Plasmidsmentioning

confidence: 99%

Recent advances in the genetics of the clostridia

Young¹

1989

FEMS Microbiology Letters

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“…The most frequently used shuttle vectors in strain ATCC 824 are derived from either Enterococcus faecalis pAM␤1 (Leblanc and Lee, 1984), C. butyricum pCBU2 (Luczak et al, 1985), or Bacillus subtilis pIM13 (Monod et al, 1986). Both replicative and non-replicative plasmids (used for insertional gene inactivation) carry genes specifying resistance to erythromycin.…”

Section: Introductionmentioning

confidence: 99%

Genetic manipulation of acid and solvent formation in Clostridium acetobutylicum ATCC 824

Green

Bennett

1998

Biotechnol. Bioeng.

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The genes coding for enzymes involved in butanol or butyrate formation were subcloned into a novel Escherichia coli-Clostridium acetobutylicum shuttle vector constructed from pIMP1 and a chloramphenicol acetyl transferase gene. The resulting replicative plasmids, referred to as pTHAAD (aldehyde/alcohol dehydrogenase) and pTHBUT (butyrate operon), were used to complement C. acetobutylicum mutant strains, in which genes encoding aldehyde/alcohol dehydrogenase (aad) or butyrate kinase (buk) had been inactivated by recombination with Em r constructs. Complementation of strain PJC4BK (buk mutant) with pTHBUT restored butyrate kinase activity and butyrate production during exponential growth. Complementation of strain PJC4AAD (aad mutant) with pTHAAD restored NAD(H)-dependent butanol dehydrogenase activity, NAD(H)-dependent butyraldehyde dehydrogenase activity and butanol production during solventogenic growth. The development of an alternative selectable marker makes it is possible to overexpress genes, via replicative plasmids, in mutant strains that lack specific enzyme activities, thereby expanding the number of possible genetic manipulations that can be performed in C. acetobutylicum.

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Construction of Clostridium butyricum hybrid plasmids and transfer to Bacillus subtilis

Cited by 5 publications

References 28 publications

Stable inheritance of shuttle vectors based on plasmid pIM13 in a mutant strain of Clostridium acetobutylicum

Stable inheritance of shuttle vectors based on plasmid pIM13 in a mutant strain of Clostridium acetobutylicum

Recent advances in the genetics of the clostridia

Genetic manipulation of acid and solvent formation in Clostridium acetobutylicum ATCC 824

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