Nuclear transformation of Volvox carteri

Stable nuclear transformation of Volvox carteri was achieved using the cloned V. carteri nitA+ gene (which encodes nitrate reductase) to complement a nitA- mutation. Following bombardment of mutant cells with plasmid-coated gold particles, putative transformants able to utilize nitrate as a nitrogen...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1994-05, Vol.91 (11), p.5080-5084
Main Authors: Schiedlmeier, B, Schmitt, R, Muller, W, Kirk, M.M, Gruber, H, Mages, W, Kirk, D.L
Format: Article
Language:English
Subjects:
Blotting, Southern
Cell Differentiation - genetics
CHLOROPHYCEAE
Chlorophyta - cytology
Chlorophyta - enzymology
Chlorophyta - genetics
DNA
Embryonic cells
Embryos
Flowers & plants
Freshwater
GENE
GENES
Genetic loci
Genetic Markers
Genetic vectors
Genetics
Genomics
Germ cells
Mutation
NITRATE REDUCTASE
Nitrate Reductases - genetics
Nitrates
NITRATO REDUCTASA
Plasmids
RECOMBINACION
RECOMBINAISON
Somatic cells
TRANSFORMACION GENETICA
TRANSFORMATION GENETIQUE
Transformation, Genetic
Volvox carteri
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Summary:Stable nuclear transformation of Volvox carteri was achieved using the cloned V. carteri nitA+ gene (which encodes nitrate reductase) to complement a nitA- mutation. Following bombardment of mutant cells with plasmid-coated gold particles, putative transformants able to utilize nitrate as a nitrogen source were recovered with an efficiency of approximately 2.5 X 10(-5). DNA analysis indicated that the plasmid integrated into the genome, often in multiple copies, at sites other than the nitA locus. Cotransformants were recovered with a frequency of 40-80% when cells were cobombarded with a selected and an unselected marker. Thus, V. carteri becomes one of the simplest multicellular organisms that is accessible to detailed molecular studies of genes regulating cellular differentiation and morphogenesis
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.91.11.5080