Linked Life Data

9304873

Source:http://linkedlifedata.com/resource/pubmed/id/9304873

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1997-10-10
pubmed:databankReference
pubmed:abstractText
A gene cloning system for Amycolatopsis mediterranei DSM5908, the producer of the glycopeptide antibiotic balhimycin, was developed for analysis of peptide synthetase genes. A modified direct transformation procedure was used to introduce DNA. The efficiency of DNA uptake depended on the age of the culture: mycelium of early stationary phase (52-55 h) cultures resulted in optimal transformation frequencies. Using the novel non-replicative integration vector pSP1, gene disruption plasmids were constructed. Highest integration frequencies were observed when the DNA was isolated from the dam/dcm Escherichia coli strain JM110. The efficiency of integration depended directly on the size of the cloned insert. Plasmids with fragments smaller than 1 kilobase (kb) were difficult to integrate. In gene replacement experiments a high double cross-over rate (31%) was demonstrated. Oligonucleotides derived from conserved regions of peptide synthetases were designed to identify balhimycin biosynthesis genes. Using these gene probes in plaque hybridization experiments, we identified peptide synthetase homologous DNA fragments in a lambda library of A. mediterranei. One peptide synthetase gene fragment was characterized by DNA sequencing and the results revealed a complete amino acid activating domain of a peptide synthetase gene, designated aps. The disruption of aps neither influenced balhimycin biosynthesis nor generated another apparent phenotype.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0168-1656
pubmed:author
pubmed:issnType
Print
pubmed:day
11
pubmed:volume
56
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
115-28
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1997
pubmed:articleTitle
Cloning and analysis of a peptide synthetase gene of the balhimycin producer Amycolatopsis mediterranei DSM5908 and development of a gene disruption/replacement system.
pubmed:affiliation
Eberhard-Karls-Universität Tübingen, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't