Linked Life Data

14586556

Source:http://linkedlifedata.com/resource/pubmed/id/14586556

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pubmed-article:14586556pubmed:abstractTextThe 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.lld:pubmed
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pubmed-article:14586556pubmed:dateRevised2006-11-15lld:pubmed
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pubmed-article:14586556pubmed:articleTitleAn improved and highly standardised transformation procedure allows efficient production of single and multiple targeted gene-knockouts in a moss, Physcomitrella patens.lld:pubmed
pubmed-article:14586556pubmed:affiliationPlant Biotechnology, Freiburg University, Schaenzlestrasse 1, 79104 Freiburg, Germany.lld:pubmed
pubmed-article:14586556pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:14586556pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed
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