Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
1993-5-20
pubmed:abstractText
A new culture method for the injection of tobacco mesophyll protoplasts has been established. The protoplasts are embedded in a thin layer of alginate and are nourished from the medium in the underlying basis layer. In the alginate layer the protoplasts regenerate to calli at a frequency of up to 80%. Embedded protoplasts can be selected either with 50 mg l-1 kanamycin or 5 mg l-1 paromomycin. Single resistant cells can be recovered from about 10,000 sensitive cells in one alginate layer. Injection of the neo gene (coding for neomycin phosphotransferase II) into protoplast derived single cells in the alginate layer results in kanamycin resistant colonies that can be regenerated to mature plants. These plants express the neomycin phosphotransferase as shown by enzyme activity assay. The integration of the transgene into the plant genome could be proved by Southern hybridization to high molecular weight DNA. With this culture method 100 cells can be injected per hour. Transformation frequencies range from 2 to 20%. In crossing experiments, it was shown that the foreign gene is transmitted to the next generation in a Mendelian fashion.
pubmed:language
eng
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:author
pubmed:geneSymbol
neo
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
23-30
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:1668908-Base Sequence, pubmed-meshheading:1668908-Blotting, Southern, pubmed-meshheading:1668908-Cells, Cultured, pubmed-meshheading:1668908-Crosses, Genetic, pubmed-meshheading:1668908-DNA, pubmed-meshheading:1668908-Genetic Techniques, pubmed-meshheading:1668908-Kanamycin Kinase, pubmed-meshheading:1668908-Kanamycin Resistance, pubmed-meshheading:1668908-Microinjections, pubmed-meshheading:1668908-Molecular Sequence Data, pubmed-meshheading:1668908-Oligodeoxyribonucleotides, pubmed-meshheading:1668908-Phosphotransferases, pubmed-meshheading:1668908-Plants, Genetically Modified, pubmed-meshheading:1668908-Plants, Toxic, pubmed-meshheading:1668908-Plasmids, pubmed-meshheading:1668908-Protoplasts, pubmed-meshheading:1668908-Tobacco, pubmed-meshheading:1668908-Transfection, pubmed-meshheading:1668908-Transformation, Genetic
pubmed:articleTitle
An improved approach for transformation of plant cells by microinjection: molecular and genetic analysis.
pubmed:affiliation
Institute for Plant Sciences, Swiss Federal Institute of Technology, Zürich, Switzerland.
pubmed:publicationType
Journal Article