Abstract:
:The analysis of the virulence determinants of phytopathogenic Rhodococcus fascians has been hampered by the lack of a system for introducing exogenous DNA. We investigated the possibility of genetic transformation of R. fascians by high-voltage electroporation of intact bacterial cells in the presence of plasmid DNA. Electrotransformation in R. fascians D188 resulted in transformation frequencies ranging from 10/mug of DNA to 10/mug of DNA, depending on the DNA concentration. The effects of different electrical parameters and composition of electroporation medium on transformation efficiency are presented. By this transformation method, a cloning vector (pRF28) for R. fascians based on an indigenous 160-kilobase (chloramphenicol and cadmium resistance-encoding) plasmid pRF2 from strain NCPPB 1675 was developed. The origin of replication and the chloramphenicol resistance gene on pRF28 were used to construct cloning vectors that are capable of replication in R. fascians and Escherichia coli. The electroporation method presented was efficient enough to allow detection of the rare integration of replication-deficient pRF28 derivatives in the R. fascians D188 genome via either homologous or illegitimate recombination.
journal_name
Appl Environ Microbioljournal_title
Applied and environmental microbiologyauthors
Desomer J,Dhaese P,Montagu MVdoi
10.1128/AEM.56.9.2818-2825.1990keywords:
subject
Has Abstractpub_date
1990-09-01 00:00:00pages
2818-25issue
9eissn
0099-2240issn
1098-5336journal_volume
56pub_type
杂志文章abstract::In a field study of 29 dairy farms, Pseudomonas aeruginosa was isolated more frequently (P = 0.05) from milking parlor udder wash water systems containing iodophor germicides than from those with no germicide. Most available iodine (AI2) concentrations were below the recommended level of 25 ppm (25 microgram/ml). Rubb...
journal_title:Applied and environmental microbiology
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:1982-03-01 00:00:00
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pub_type: 杂志文章
doi:10.1128/AEM.49.3.664-666.1985
更新日期:1985-03-01 00:00:00
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pub_type: 杂志文章
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
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更新日期:2014-09-01 00:00:00
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pub_type: 杂志文章
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更新日期:2002-12-01 00:00:00
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journal_title:Applied and environmental microbiology
pub_type: 杂志文章
doi:10.1128/AEM.57.3.889-891.1991
更新日期:1991-03-01 00:00:00