Linked Life Data

9097047

Source:http://linkedlifedata.com/resource/pubmed/id/9097047

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1997-6-17
pubmed:abstractText
We have previously constructed a cloning/sequencing vector, with an in vivo system capable of creating nested deletions from the end of transposon Tn3, which is useful for sequencing large DNAs. Here we report an in vitro system which uses an ammonium sulfate fraction of extract from E. coli cells harboring a Tn3 transposase overproducer plasmid to generate nested deletions. A key feature of the procedure is exhaustive digestion of the reaction products with a restriction enzyme that cleaves only between the Tn3 "right" terminus and the cloned fragment. This step reduces the noise level due to mechanisms other than deletions from the Tn3 terminus, and facilitates detection and isolation of the desired deletion products. This system enables us to save at least 2 days' time when obtaining the necessary deletions compared with the in vivo system.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1340-2838
pubmed:author
pubmed:issnType
Print
pubmed:day
31
pubmed:volume
3
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
431-3
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed:year
1996
pubmed:articleTitle
Nested deletions from a fixed site as an aid to nucleotide sequencing: an in vitro system using Tn3 transposase.
pubmed:affiliation
Laboratory of Molecular Biology, Kansai Medical University, Osaka, Japan. morita@makino.kmu.ac.jp
pubmed:publicationType
Journal Article