Transfer of Tn1545 and Tn916 to Clostridium acetobutylicum

Woolley, R.C.; Pennock, A.; Ashton, R.; Davies, A.M.C.; Young, Michael

doi:10.1016/0147-619x(89)90027-9

Cited by 55 publications

(25 citation statements)

References 32 publications

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“…Tn1545 (Courvalin & Carlier, 1986) was transferred from Enterococcus faecalis BM4110::Tn1545 to C. acetobutylicum NCIMB 8052 as described by Woolley et al (1989).…”

Section: S R Wilkinson and M Youngmentioning

confidence: 99%

“…This was demonstrated using Tn1545, a conjugative transposon that is readily transferred from Ent. faecalis to C. acetobutylicum NCIMB 8052 (Woolley et al, 1989). This large (25.3 kbp) element lacks restriction sites for both SmaI and ApaI (Caillaud et al, 1987a) and therefore, DNA fragments into which it had inserted increased in size by about 25 kbp.…”

Section: Estimation Of Genome Sizesmentioning

confidence: 99%

“…These data, considered in conjunction with results obtained previously from comparative studies of physiology, biochemistry, cell wall lytic enzymes, susceptibility to the Ent. faecalis conjugative transposon Tn916, and nucleic acid hybridization (Woolley, 1988 ;Woolley et al, 1989;Woolley & Morris, 1990;Croux & Garcia, 1991), indicate that the various strains denoted C . acetobutylicum do not form a homogeneous species.…”

Section: Estimation Of Genome Sizesmentioning

confidence: 99%

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Wide diversity of genome size among different strains of Clostridium acetobutylicum

Wilkinson

Young

1993

Journal of General Microbiology

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The sizes of the genomes of five strains of Clostridium acetobutylicum were estimated by summing the sizes of macro-restriction fragments produced after cleavage of their DNA with SmaI and ApaI. The five C. acetobutylicum strains fell into three discrete groups. The NCIMB 8052 and NI-4081 strains have 6.5 Mbp genomes and restriction profiles of their DNA were indistinguishable. The sizes of the genomes of the ATCC 824 and DSM 1731 strains were about 4-0 and 3.5 Mbp (minimum values), respectively, and the restriction profiles of their DNA were very similar. One notable difference was the presence of an additional 532 kbp SmaI fragment in the DNA of strain ATCC 824. The NCP 262 strain of C. acetobutylicum has a genome size of 2-85 Mbp. Restriction fragment profiles of DNA from strains belonging to different groups were not obviously similar. Probes made from several cloned ATCC 824 genes hybridized with DNA fragments of different sizes from strains representative of each group. These data add further weight to the considerable body of evidence indicating that the species C. acetobutylicum comprises a heterogeneous collection of strains.

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“…Tn1545 (Courvalin & Carlier, 1986) was transferred from Enterococcus faecalis BM4110::Tn1545 to C. acetobutylicum NCIMB 8052 as described by Woolley et al (1989).…”

Section: S R Wilkinson and M Youngmentioning

confidence: 99%

Section: Estimation Of Genome Sizesmentioning

confidence: 99%

Section: Estimation Of Genome Sizesmentioning

confidence: 99%

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Wide diversity of genome size among different strains of Clostridium acetobutylicum

Wilkinson

Young

1993

Journal of General Microbiology

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“…They may be introduced by either electroporation (46) or intergeneric conjugation with Escherichia coli donors (56). This strain is also susceptible to Tn1545 mutagenesis (58). Homologous recombination between incoming sequences and their counterparts in the bacterial chromosome has recently been demonstrated (55).…”

mentioning

confidence: 99%

“…Conjugative transfer of Tn1545. Strains of C. beijerinckii harboring Tn1545 were isolated after the wild-type NCIMB 8052 strain had been conjugated with an Enterococcus faecalis BM4110::Tn1545 donor, as previously described (58).…”

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confidence: 99%

Physical map of the Clostridium beijerinckii (formerly Clostridium acetobutylicum) NCIMB 8052 chromosome

Wilkinson

Young

1995

J Bacteriol

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A combined physical and genetic map of the single, circular, 6.7-Mbp chromosome of the NCIMB 8052 strain of Clostridium beijerinckii (formerly Clostridium acetobutylicum) has been constructed by using a combination of cloned DNA fragments as hybridization probes and a bank of strains harboring insertions of the conjugative transposon Tn1545. The positions of 81 restriction endonuclease cleavage sites and 32 genes have been determined. Eight genes concerned with solventogenic fermentation are found at three different locations. The chromosome contains at least 13 rrn operons, 11 of which have been located on the map. Their transcriptional orientation diverges from the presumed location of the replication origin.Clostridium beijerinckii (formerly Clostridium acetobutylicum) is an obligately anaerobic spore-forming bacterium that produces acetone, butanol, and small amounts of ethanol when grown on sugary substrates. This process, known as ABE fermentation, has been used extensively for the industrial scale production of acetone and butanol (28). Several different strains are currently under investigation in various laboratories worldwide. They have been separated into three distinct groups on the basis of their biochemical and physiological characteristics, the extent of hybridization between their DNAs, and macrorestriction fragment profiles of their genomes (54, 57). The NCIMB 8052 strain, which belongs to the group that has the largest chromosome, is amenable to genetic manipulation (59, 60). It lacks the active restriction system that is found in the ATCC 824 strain (38). The NCIMB 8052 strain has no indigenous plasmids, but it supports replication of a variety of vectors based on plasmids from Clostridium butyricum, Enterococcus faecalis, and Streptococcus cremoris (41). They may be introduced by either electroporation (46) or intergeneric conjugation with Escherichia coli donors (56). This strain is also susceptible to Tn1545 mutagenesis (58). Homologous recombination between incoming sequences and their counterparts in the bacterial chromosome has recently been demonstrated (55).As an additional aid to genetic analysis of this organism, we have constructed a physical map of the 6.7-Mbp chromosome using pulsed-field gel electrophoresis (PFGE) (50). The application of PFGE to the analysis of bacterial genomes has revealed that the majority of bacteria have single circular chromosomes. However, several exceptions to this general rule have been described. Some bacteria, such as Borrelia burgdorferi and Rhodococcus fascians, have linear chromosomes (4,20,24), whereas Streptomyces lividans and Streptomyces coelicolor have linear chromosomes that may become circular as a result of deletions spanning both telomeres (34,35). Still others, such as Brucella melitensis and Leptospira interrogans, have segmented genomes (40, 63). The segmented genome of Agrobacterium tumefaciens has one circular and one linear component (1). The results reported here indicate that the NCIMB 8052 strain of C. beijerinckii, like most othe...

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Clostridia

Bahl¹,

Dürre²

1993

Biotechnology

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Transfer of Tn1545 and Tn916 to Clostridium acetobutylicum

Cited by 55 publications

References 32 publications

Wide diversity of genome size among different strains of Clostridium acetobutylicum

Wide diversity of genome size among different strains of Clostridium acetobutylicum

Physical map of the Clostridium beijerinckii (formerly Clostridium acetobutylicum) NCIMB 8052 chromosome

Clostridia

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