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Hemp (Cannabis sativa L.)
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Feeney, Mistianne and Punja, Zamir K. (2007) Hemp (Cannabis sativa L.). In: Wang, Kan, (ed.) Agrobacterium Protocols Volume 2. Methods in Molecular Biology (Part VII). Humana Press, pp. 373-382. ISBN 9781588298430
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Official URL: http://dx.doi.org/10.1385/1-59745-131-2:373
Abstract
Hemp (Cannabis sativa L.) suspension culture cells were transformed with Agrobacterium tumefaciens strain EHA101 carrying the binary plasmid pNOV3635. The plasmid contains a phosphomannose isomerase (PMI) selectable marker gene. Cells transformed with PMI are capable of metabolizing the selective agent, mannose, whereas cells not expressing the gene are incapable of using the carbon source and will stop growing. Callus masses proliferating on selection were screened for PMI expression using a chlorophenol red assay. Genomic DNA was extracted from putatively transformed callus lines and the presence of the PMI gene was confirmed using polymerase chain reaction and Southern hybridization. Using this method, an average transformation frequency of 31.23% ± 0.14 was obtained for all transformation experiments, with a range of 15.1 to 55.3%.
Item Type: | Book Item | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Series Name: | Methods in Molecular Biology | ||||
Publisher: | Humana Press | ||||
ISBN: | 9781588298430 | ||||
Book Title: | Agrobacterium Protocols Volume 2 | ||||
Editor: | Wang, Kan | ||||
Official Date: | 2007 | ||||
Dates: |
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Number: | Part VII | ||||
Page Range: | pp. 373-382 | ||||
DOI: | 10.1385/1-59745-131-2:373 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
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