Cloning of the YenI restriction endonuclease and methyltransferase from Yersinia enterocolitica serotype O8 and construction of a transformable R−M+ mutant

Kinder, Susan A.; Badger, Julie L.; Bryant, Gene O.; Pepe, Jeffrey C.; Miller, Virginia L.

doi:10.1016/0378-1119(93)90478-l

Cited by 169 publications

(96 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mutant strains carrying an insertion mutation that inactivated crp or cyaA were generated using the suicide plasmid pEP185.2 as previously described (15). DNA fragments corresponding to an internal region of the target gene were generated by PCR with the following oligonucleotide primers: for crp, YP-CRP1 (5Ј-GTAAGCCA CAAACAGACCCG-3Ј) and YP-CRP2 (5Ј-CCTGAATCAGTTGACGGAAC-3Ј); and for cyaA, Pestis cya 1 (5Ј-CAGATCTGAGTCTGCTACCGATCC-3Ј) and Pestis cya 2 (5Ј-CAGATCTGTCAACCAGGAAGAAGC-3Ј).…”

Section: Methodsmentioning

confidence: 99%

Direct Transcriptional Control of the Plasminogen Activator Gene of Yersinia pestis by the Cyclic AMP Receptor Protein

Kim¹,

Chauhan²,

Motin

et al. 2007

J Bacteriol

View full text Add to dashboard Cite

Horizontal gene transfer events followed by proper regulatory integration of a gene drive rapid evolution of bacterial pathogens. A key event in the evolution of the highly virulent plague bacterium Yersinia pestis was the acquisition of plasmid pPCP1, which carries the plasminogen activator gene, pla. This promoted the bubonic form of the disease by increasing bacterial dissemination from flea bite sites and incidentally enhanced replication in respiratory airways during pneumonic infection. We determined that expression of pla is controlled by the global regulator cyclic AMP (

show abstract

Section: Methodsmentioning

confidence: 99%

Direct Transcriptional Control of the Plasminogen Activator Gene of Yersinia pestis by the Cyclic AMP Receptor Protein

Kim¹,

Chauhan²,

Motin

et al. 2007

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…1A), suggesting that the plasmids were unrelated. However, Southern hybridization studies (15) indicated that pFN1 and pFN2 share homology with each other but not with pFN3 ( Fig. 1B and C).…”

mentioning

confidence: 96%

Native Plasmids of Fusobacterium nucleatum : Characterization and Use in Development of Genetic Systems

Haake

Yoder²,

Attarian³

et al. 2000

J Bacteriol

View full text Add to dashboard Cite

Three native plasmids of Fusobacterium nucleatum were characterized, including DNA sequence analysis of one plasmid, pFN1. A shuttle plasmid, pHS17, capable of transforming Escherichia coli and F. nucleatum ATCC 10953 was constructed with pFN1. pHS17 was stably maintained in the F. nucleatum transformants, and differences in the transformation efficiencies suggested the presence of a restriction-modification system in F. nucleatum.

show abstract

“…Our analysis of the ATCC 10953 sequence indicates the existence of at least two putative restriction endonuclease genes (unpublished data). Strategies to pre-methylate vector DNA (12), or to knockout restriction endonucleases in strains amenable to transformation (25), may facilitate further advances in gene transfer systems.…”

Section: Discussionmentioning

confidence: 99%

“…The forward primer, PA1, was specific to sequence in the rnr gene located upstream of the rnr gene fragment used in the integration plasmid; the reverse primer was specific to the erythromycin resistance cassette present in the integration plasmid. For Southern blot analysis, chromosomal DNA was digested with AccI, subjected to gel electrophoresis and transferred to nitrocellulose, then probed with a PCR-amplified 32 P-radiolabelled rnr gene probe, as previously described (23,25). PCR primers used for amplification of the rnr probe, which corresponded to bp 170 to 1842 of the rnr gene, were the SA1 and SA2.…”

Section: Targeted Integration Mutagenesismentioning

confidence: 99%

Efficient gene transfer and targeted mutagenesis in Fusobacterium nucleatum

Haake

Yoder

Gerardo

2006

Plasmid

View full text Add to dashboard Cite

Fusobacterium nucleatum is a Gram-negative anaerobe important in dental biofilm ecology and infectious diseases with significant societal impact. The lack of efficient genetic systems has hampered molecular analyses in this microorganism. We previously reported construction of a shuttle plasmid, pHS17, using the native fusobacterial plasmid pFN1 and an erythromycin resistance cassette. However, the host range of pHS17 was restricted to F. nucleatum, ATCC 10953 and the transformation efficiency was limited. This study was undertaken to improve genetic systems for molecular analysis in F. nucleatum. We identified a second F. nucleatum strain, ATCC 23726, which is transformed with improved efficiency compared to ATCC 10953. Two novel second generation pFN1-based shuttle plasmids, pHS23 and pHS30, were developed and enable transformation of ATCC 23726 at 6.2 x 10 4 and 1.5 x 10 6 transformants/microgram of plasmid DNA, respectively. The transformation efficiency of pHS30, which harbors a catP gene conferring resistance to chloramphenicol, was more than 1,000-fold greater than that of pHS17. The improved transformation efficiency facilitated disruption of the chromosomal rnr gene using a suicide plasmid pHS19, the first demonstration of targeted mutagenesis in F. nucleatum. These results provide significant advances in the development of systems for molecular analysis in F. nucleatum.

show abstract

Cloning of the YenI restriction endonuclease and methyltransferase from Yersinia enterocolitica serotype O8 and construction of a transformable R−M+ mutant

Cited by 169 publications

References 26 publications

Direct Transcriptional Control of the Plasminogen Activator Gene of Yersinia pestis by the Cyclic AMP Receptor Protein

Direct Transcriptional Control of the Plasminogen Activator Gene of Yersinia pestis by the Cyclic AMP Receptor Protein

Native Plasmids of Fusobacterium nucleatum : Characterization and Use in Development of Genetic Systems

Efficient gene transfer and targeted mutagenesis in Fusobacterium nucleatum

Contact Info

Product

Resources

About