Transformation of Clostridium perfringens L forms with shuttle plasmid DNA

Mahony, D. E.; Mader, J A; Dubel, J.

doi:10.1128/aem.54.1.264-267.1988

Cited by 21 publications

(11 citation statements)

References 10 publications

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“…We found that cultures of L-13 in BHIS survived only 2 to 3 days at 37°C or room temperature, 4 to 5 days at 4°C, and 3 to 4 weeks at 4°C in BHIS with 0.25% agar (the semisolid medium described by Mahony et al [7]). Chopped-meat cultures (4) of L-13 were prepared by withdrawing 5 ml of the Trypticase liquid medium from a chopped-meat tube and replacing the liquid with 5 ml of an overnight culture of L-13 in BHIS.…”

mentioning

confidence: 81%

“…This paper describes the construction and performance of a shuttle plasmid (pHR106) which contains a chloramphenicol resistance marker gene and a set of unique cloning sites and which transforms L-phase C. perfringens L-13 with ease and reliability. The accompanying paper by Mahony et al (7) presents the background of strain L-13 and data on its transformation.…”

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confidence: 99%

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Development of a new shuttle plasmid system for Escherichia coli and Clostridium perfringens

Roberts

Holmes

Hylemon

1988

Appl Environ Microbiol

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We constructed a 7.9-kilobase-pair recombinant shuttle plasmid, designated pHR106, by combining desired segments of three plasmids: an Escherichia coli plasmid (pSL100) which provides a multiple cloning site, a Clostridium perfringens plasmid (pJU122) which provides a clostridial origin of replication, and an E. coli plasmid (pJIR62) which provides an E. coli origin of replication, an ampicillin resistance gene, and a chloramphenicol resistance gene of clostridial origin. The shuttle plasmid transformed E. coli HB101 with a frequency of 1 transformant per 104 viable cells and C. perfringens L-phase strain L-13 with a frequency of approximately 1 transformant per 106 viable cells. Because of the set of unique cloning sites and the chloramphenicol resistance marker, this shuttle plasmid should be particularly useful for studies of gene regulation and for enzyme production with C. perfringens.

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confidence: 81%

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confidence: 99%

Development of a new shuttle plasmid system for Escherichia coli and Clostridium perfringens

Roberts

Holmes

Hylemon

1988

Appl Environ Microbiol

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“…Recently, there has been a renewed interest in L-form bacteria. This has arisen because of their apparent ability to associate with plant cells (Jones & Paton 1973;Aloysius & Paton 1984;Paton 1987) and in the case of Clostridium perfringens, the increased ease of their genetic transformation compared with the cell-walled form (Mahony et al 1988).…”

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confidence: 99%

Induction and cultivation of a stable L‐form of Bacillus subtilis

Allan

1991

Journal of Applied Bacteriology

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The induction of L-forms of Bacillus subtilis from protoplasts is described. The method involved the frequent subculture of the unstable L-form on a growth medium supplemented with lysozyme and horse serum. A stable culture, which did not revert when lysozyme and horse serum were omitted from the medium, was obtained after 13 subcultures. This culture could be grown on solid and in liquid medium by routine microbiological methods. Long-term storage of these cells was achieved by freeze drying and maintenance in glycerol at -70 degrees C. The cultural adaptability of the L-form is described and discussed with respect to methods of cultivation and growth.

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“…Squires et al (19) constructed shuttle vectors and demonstrated their transferability into Escherichia coli and L-phase variants of C. perfringens. C. perfringens L-phase variant transformation was described by Heefner et al (9) and, more recently, Mahony et al (12). Roberts et al (16) recently described an E. coli-C. perfringens shuttle vector (pHR106) which was transformed into L-phase variants of C. perfringens.…”

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Construction of an Escherichia coli-Clostridium perfringens shuttle vector and plasmid transformation of Clostridium perfringens

Kim

Blaschek

1989

Appl Environ Microbiol

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A stable shuttle vector which replicates in Escherichia coli and Clostridium perfringens was constructed by ligating a 3.6-kilobase (kb) fragment of plasmid pBR322 with C. perfringens plasmid plB101 (3.1 kb). The marker for this shuttle plasmid originated from the 1.3-kb chloramphenicol resistance gene of plasmid pHR106. The resulting shuttle vector, designated pAK201, is 8 kb in size and codes for resistance to 20 ,ug of chloramphenicol per ml in both E. coli and C. perfringens. Following shuttle vector construction in E. coli, plasmid pAK201 was transformed into E. coli HB101 and C. perfringens ATCC 3624A, using intact cell electroporation. The transformation frequencies were 106 and 104 transformants per ,ug of DNA in E. coli and C. perfringens, respectively. Restriction enzyme analysis of the chimera isolated from transformants of both microorganisms suggested that the plasmids were identical. Reciprocal transformation experiments in E. coli and C. perfringens indicated no difference in transformation frequency. Plasmid pAK201 was stable in C. perfringens following repeated transfer in the absence of chloramphenicol pressure. The restriction map of plasmid pAK201 shows six unique cut sites which should be useful for future genetic analysis and C. perfringens gene library construction.

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Transformation of Clostridium perfringens L forms with shuttle plasmid DNA

Cited by 21 publications

References 10 publications

Development of a new shuttle plasmid system for Escherichia coli and Clostridium perfringens

Development of a new shuttle plasmid system for Escherichia coli and Clostridium perfringens

Induction and cultivation of a stable L‐form of Bacillus subtilis

Construction of an Escherichia coli-Clostridium perfringens shuttle vector and plasmid transformation of Clostridium perfringens

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