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  Identification of the fire blight pathogen, Erwinia amylovora, by PCR assays with chromosomal DNA

Bereswill, S., Bugert, P., Bruchmüller, I., & Geider, K. (1995). Identification of the fire blight pathogen, Erwinia amylovora, by PCR assays with chromosomal DNA. Applied and Environmental Microbiology, 61(7), 2636-2642. Retrieved from http://www.ncbi.nlm.nih.gov/pmc/articles/PMC167536/.

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Genre: Journal Article
Alternative Title : Identification of the fire blight pathogen, Erwinia amylovora, by PCR assays with chromosomal DNA

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ApplEnvironMicrobiol_61_1995_2636.pdf (Any fulltext), 254KB
 
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 Creators:
Bereswill, Stefan1, Author              
Bugert, Peter1, Author              
Bruchmüller, Iris2, Author              
Geider, Klaus1, Author              
Affiliations:
1Research Group Prof. Dr. Geider, Max Planck Institute for Medical Research, Max Planck Society, ou_1497738              
2Max Planck Institute for Medical Research, Max Planck Society, ou_1125545              

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 Abstract: Erwinia amylovora, the causative agent of fire blight, was identified independently from the common plasmid pEA29 by three different PCR assays with chromosomal DNA. PCR with two primers was performed with isolated DNA and with whole cells, which were directly added to the assay mixture. The oligonucleotide primers were derived from the ams region, and the PCR product comprised the amsB gene, which is involved in exopolysaccharide synthesis. The amplified fragment of 1.6 kb was analyzed, and the sequence was found to be identical for two E. amylovora strains. The identity of the PCR products was further confirmed by restriction analysis. The 1.6-kb signal was also used for detection of the fire blight pathogen in the presence of other plant-associated bacteria and in infected plant tissue. For further identification of isolated strains, the 16S rRNA gene of E. amylovora and other plant-associated bacteria was amplified and the products were digested with the restriction enzyme HaeIII. The pattern obtained for E. amylovora was different from that of other bacteria. The sequence of the 16S rRNA gene was determined from a cloned fragment and was found to be closely related to the sequences of Escherichia coli and other Erwinia species. Finally, arbitrarily primed PCR with a 17-mer oligonucleotide derived from the sequence of transposon Tn5 produced a unique banding pattern for all E. amylovora strains investigated. These methods expand identification methods for E. amylovora, which include DNA hybridization and a PCR technique based on plasmid pEA29.

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Language(s): eng - English
 Dates: 1995-07-01
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 665207
URI: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC167536/
Other: 6871
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Title: Applied and Environmental Microbiology
  Other : Appl. Environ. Microbiol.
Source Genre: Journal
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Publ. Info: American Society for Microbiology (ASM)
Pages: - Volume / Issue: 61 (7) Sequence Number: - Start / End Page: 2636 - 2642 Identifier: ISSN: 0099-2240
CoNE: https://pure.mpg.de/cone/journals/resource/954927519600