Transformation of cucumber tissues by microprojectile bombardment: identification of plants containing functional and non-functional transferred genes.
The microprojectile bombardment method was used to transfer DNA into embryogenic callus of cucumber (Cucumis sativus), and stably transformed cucumber plant lines were obtained. A total of 107 independently regenerated cucumber plants were assayed for the presence and expression of the transferred Nos-NPTII gene (encoding nopaline synthase-neomycin phosphotransferase II). Genomic blot hybridization analyses showed that a high percentage (16%) of the cucumber plants were transformed with Nos-NPTII; however, only about 25% of these transgenic plants expressed Nos-NPTII. Inactivity of Nos-NPTII in many of the transformed cucumber plants may be associated with the transfer of multiple copies of Nos-NPTII. PCR and genomic blot hybridization analyses were used to show that the transferred gene was inherited in the subsequent plant generation.
Chee
P P
PP
Molecular Biology Research Unit, Upjohn Company, Kalamazoo, MI 49007.
Slightom
J L
JL
D000594:Amino Acid Oxidoreductases / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods*; D019868:Kanamycin Kinase; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D010770:Phosphotransferases / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction; D014162:Transfection / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase; D010770:Phosphotransferases; D019868:Kanamycin Kinase