title: Hemp (Cannabis sativa L.).
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%.
authors.affiliation: School of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom.
authors.forename: Mistianne
authors.initials: M
authors.lastname: Feeney
authors.forename: Zamir K
authors.initials: ZK
authors.lastname: Punja
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000254:growth & development; D002188:Cannabis / Q000166:cytology / Q000235:genetics* / Q000254:growth & development*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D005818:Genetic Engineering / Q000379:methods*; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
chemical_list: D018744:DNA, Plant; D008359:Mannose-6-Phosphate Isomerase