Isolation of a single-stranded plasmid from Clostridium acetobutylicum NCIB 6444

Kim, A. Y.; Vertès, Alain A.; Blaschek, Hans P.

doi:10.1128/aem.56.6.1725-1728.1990

Cited by 12 publications

(3 citation statements)

References 19 publications

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“…C. acetobutylicum ATCC 824 was obtained from the American Type Culture Collection. For DNA preparation, C. acetobutylicum was grown under anaerobic conditions in batch mode at 37°C on trypticase–glucose–yeast extract medium (2YTG)[7]. For Northern (RNA) experiments and amylolytic assays, continuous cultures (dilution rate of 0.05 h −1 ) were run at 35°C in phosphate‐limited synthetic medium as previously described[8].…”

Section: Methodsmentioning

confidence: 99%

amyP, a reporter gene to study strain degeneration inClostridium acetobutylicumATCC 824

Sabathé

Croux

Cornillot

et al. 2002

FEMS Microbiology Letters

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Clostridium acetobutylicum produces an extracellular alpha-amylase when grown on glucose as the sole carbon source. This enzyme was previously characterized from a biochemical point of view but its encoding gene was never identified. The 2283-bp amyP gene encodes a 83013-Da mature protein with an N-terminal domain that exhibits strong identity to the family 13 glycosyl hydrolases such as the Bacillus alpha-amylases. Transcriptional analysis revealed that amyP is transcribed in solventogenic but not in acidogenic chemostat cultures. These results are in agreement with the extracellular alpha-amylase activities indicating that the expression of amyP is regulated at the transcriptional level. amyP is located on the pSOL1 megaplasmid that carries all the genes involved in the final steps of solvent formation. Degeneration of C. acetobutylicum has been associated to the loss of pSOL1. We demonstrate here that amyP can be used as a reporter system to quantitatively follow this phenomenon.

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Section: Methodsmentioning

confidence: 99%

amyP, a reporter gene to study strain degeneration inClostridium acetobutylicumATCC 824

Sabathé

Croux

Cornillot

et al. 2002

FEMS Microbiology Letters

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show abstract

“…acetobutylicum ATCC 824 was obtained from the American Type Culture Collection. For DNA preparation, C. acetobutylicum was grown under anaerobic con-ditions in batch mode at 37 ‡C on trypticase^glucose^yeast extract medium (2YTG) [7]. For Northern (RNA) experiments and amylolytic assays, continuous cultures (dilution rate of 0.05 h 31 ) were run at 35 ‡C in phosphate-limited synthetic medium as previously described [8].…”

Section: Organisms and Culture Conditionsmentioning

confidence: 99%

amyP, a reporter gene to study strain degeneration in Clostridium acetobutylicum ATCC 824

Sabathé¹

2002

FEMS Microbiology Letters

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Clostridium acetobutylicum produces an extracellular K-amylase when grown on glucose as the sole carbon source. This enzyme was previously characterized from a biochemical point of view but its encoding gene was never identified. The 2283-bp amyP gene encodes a 83 013-Da mature protein with an N-terminal domain that exhibits strong identity to the family 13 glycosyl hydrolases such as the Bacillus K-amylases. Transcriptional analysis revealed that amyP is transcribed in solventogenic but not in acidogenic chemostat cultures. These results are in agreement with the extracellular K-amylase activities indicating that the expression of amyP is regulated at the transcriptional level. amyP is located on the pSOL1 megaplasmid that carries all the genes involved in the final steps of solvent formation. Degeneration of C. acetobutylicum has been associated to the loss of pSOL1. We demonstrate here that amyP can be used as a reporter system to quantitatively follow this phenomenon. ß

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“…During the course of the construction of a versatile E. coli-C, acetobutylicum shuttle vector, we isolated and characterized a single-stranded plasmid from C. acetobutylicum NCIB 6444 [13]. This strain was found to continuously extrude single-stranded DNA during cell growth, an event normally only seen in filamentous phage-infected Gram-negative microorganisms [10,14].…”

Section: Phage-based Vectormentioning

confidence: 99%

Genetic systems development in the clostridia

Blaschek

1995

FEMS Microbiology Reviews

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This review describes recent developments in the genetic manipulation of the solventogenic clostridia, Clostridium acetobutylicum and C. beijerinckii. It is to be noted that our laboratory stock of C. acetobutylicum ATCC 824, which was obtained from the American Type Culture Collection, has recently been re-identified as C. beijerinckii NCIMB 8052 based on DNA similarity studies using the S1 nuclease method (personal communication, Dr. Jiann-Shin Chen, Virginia Polytechnic Institute and State University). Reference to our laboratory 824 culture has been changed to C. beijerinckii NCIMB 8052 throughout this paper in order to be consistent with this finding. The focus of this review specifically involves the characterization of an M13-1ike genetic system for the clostridia based on the pCAK1 phagemid, as well as preliminary work on development of a plasmid-based vector based on the indigenous pDM11 plasmid recovered from C. acetobutylicum NCIB 6443. The construction of a C. beijerinckii strain with amplified endoglucanase activity was achieved by inserting the engB gene from C. cellulovorans into C. beijerinckii. The successful expression of a heterologous engB gene from C. cellulovorans in C. beijerinckii NCIMB 8052 has important industrial significance for the eventual utilization of cellulose by this acetone-butanol-ethanol fermentation microorganism.

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Isolation of a single-stranded plasmid from Clostridium acetobutylicum NCIB 6444

Cited by 12 publications

References 19 publications

amyP, a reporter gene to study strain degeneration inClostridium acetobutylicumATCC 824

amyP, a reporter gene to study strain degeneration inClostridium acetobutylicumATCC 824

amyP, a reporter gene to study strain degeneration in Clostridium acetobutylicum ATCC 824

Genetic systems development in the clostridia

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