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Hemp (Cannabis sativa L.)

 Last Update: 2021-03-12
 Source: Internet
 Author: User

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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%.

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