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Hemp (Cannabis sativa L.)
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Feeney, Mistianne and Punja, Zamir K. (2015) Hemp (Cannabis sativa L.). In: Wang, Kan, (ed.) Agrobacterium Protocols Volume 2. Methods in Molecular Biology, 2 (1224). New York: Springer, pp. 319-329. ISBN 9781493916573
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Official URL: http://dx.doi.org/10.1007/978-1-4939-1658-0_25
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 medium 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 PCR 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 15.1–55.3 %.
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: | Springer | ||||
Place of Publication: | New York | ||||
ISBN: | 9781493916573 | ||||
Book Title: | Agrobacterium Protocols Volume 2 | ||||
Editor: | Wang, Kan | ||||
Official Date: | 2015 | ||||
Dates: |
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Volume: | 2 | ||||
Number: | 1224 | ||||
Page Range: | pp. 319-329 | ||||
DOI: | 10.1007/978-1-4939-1658-0_25 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published |
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