Source:http://linkedlifedata.com/resource/pubmed/id/14586556
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
6
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pubmed:dateCreated |
2003-12-19
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pubmed:abstractText |
The moss Physcomitrella patens is the only land plant known to date with highly efficient homologous recombination in its nuclear DNA, making it a unique model for plant functional genomics approaches. For high-throughput production of knockout plants, a robust transformation system based on polyethylene glycol-mediated transfection of protoplasts was developed and optimised. Both the DNA conformation and pre-culture of plants used for protoplast isolation significantly affected transformation efficiencies. Employing a newly developed PCR high-throughput method, the gene-targeting efficiency in more than 1000 plants transformed with different cDNA-based knockout constructs was determined and analysed with regard to the length and intron/exon structure of the homologous gene locus. Different targeting constructs, each containing an identical selectable marker gene, were applied as batch DNA in a single transformation experiment and resulted in double-knockout plants. Thus, the fast and efficient generation of multiple targeted gene-knockouts is now feasible in Physcomitrella.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0172-8083
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
44
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
339-47
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:14586556-Amino Acid Sequence,
pubmed-meshheading:14586556-Base Sequence,
pubmed-meshheading:14586556-Bryopsida,
pubmed-meshheading:14586556-DNA, Complementary,
pubmed-meshheading:14586556-Gene Targeting,
pubmed-meshheading:14586556-Genes, Plant,
pubmed-meshheading:14586556-Genetic Vectors,
pubmed-meshheading:14586556-Genome, Plant,
pubmed-meshheading:14586556-Green Fluorescent Proteins,
pubmed-meshheading:14586556-Luminescent Proteins,
pubmed-meshheading:14586556-Models, Genetic,
pubmed-meshheading:14586556-Plants, Genetically Modified,
pubmed-meshheading:14586556-Protoplasts,
pubmed-meshheading:14586556-Recombination, Genetic,
pubmed-meshheading:14586556-Sequence Homology, Nucleic Acid,
pubmed-meshheading:14586556-Transformation, Genetic
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pubmed:year |
2004
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pubmed:articleTitle |
An improved and highly standardised transformation procedure allows efficient production of single and multiple targeted gene-knockouts in a moss, Physcomitrella patens.
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pubmed:affiliation |
Plant Biotechnology, Freiburg University, Schaenzlestrasse 1, 79104 Freiburg, Germany.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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