The NPTII gene has been successfully transferred to the seed embryo cells of two rice varieties (Oryza sativa L. subsp. indica cv. Sanerai and Qryza sativa L. subsp. japonica cv. Nonghu No. 6) by means of electroinjec...The NPTII gene has been successfully transferred to the seed embryo cells of two rice varieties (Oryza sativa L. subsp. indica cv. Sanerai and Qryza sativa L. subsp. japonica cv. Nonghu No. 6) by means of electroinjection. Resistant calli were screened out on MS medium with 100 μg/ml Km. Transgenic rice plants were regenerated via somatic embryogenesis. Both NPTII detection and Southern blot hybridization demonstrate that the foreign gene has integrated and expressed stably in the transformants.展开更多
Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefa- ciens strain LBA4404 that harbored the binary vector pBI121 containing genes for fl-glucuronidase （GU...Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefa- ciens strain LBA4404 that harbored the binary vector pBI121 containing genes for fl-glucuronidase （GUS） and neomycin phos- photransterase （npt II）. S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets fi＇om the explants. The transformed plants resembled their parents in morphology.展开更多
The possible horizomal transfer of transgenes is of great concern when the transgenic plants are released imo the field. To test the possible transfer of nptII of transgenic trees into soil bacteria, we have used a st...The possible horizomal transfer of transgenes is of great concern when the transgenic plants are released imo the field. To test the possible transfer of nptII of transgenic trees into soil bacteria, we have used a stool DNA preparation kit to isolate the DNA from the soils in the rhizospheres of two non- and eight transgenic Eucalyptus camaldulensis trees. All the samples have provided the corresponding PCR products in the amplification with bacterial 16S RNA specific sequences, which indicates that the quality of the isolated DNA is adequate for amplification. The nptⅡ specific band has been amplified in three soil samples from the transgenic trees and even treated with filtration before the DNA isolation. This indicates that nptII DNA exists in the soil, although it is still unclear whether the DNA was in the soil particles, in the soil bacteria or in the Agrobacterium comamination which was used for the E. camaldulensis transformation. Two approaches on isolation of bacterial DNA have been suggested for testing the possibility of this event in the future.展开更多
基金Project supported by the Chinese National "7.5" Program and the Rockefeller Foundation's International Program on Rice Biotechnology.
文摘The NPTII gene has been successfully transferred to the seed embryo cells of two rice varieties (Oryza sativa L. subsp. indica cv. Sanerai and Qryza sativa L. subsp. japonica cv. Nonghu No. 6) by means of electroinjection. Resistant calli were screened out on MS medium with 100 μg/ml Km. Transgenic rice plants were regenerated via somatic embryogenesis. Both NPTII detection and Southern blot hybridization demonstrate that the foreign gene has integrated and expressed stably in the transformants.
文摘Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefa- ciens strain LBA4404 that harbored the binary vector pBI121 containing genes for fl-glucuronidase （GUS） and neomycin phos- photransterase （npt II）. S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets fi＇om the explants. The transformed plants resembled their parents in morphology.
文摘The possible horizomal transfer of transgenes is of great concern when the transgenic plants are released imo the field. To test the possible transfer of nptII of transgenic trees into soil bacteria, we have used a stool DNA preparation kit to isolate the DNA from the soils in the rhizospheres of two non- and eight transgenic Eucalyptus camaldulensis trees. All the samples have provided the corresponding PCR products in the amplification with bacterial 16S RNA specific sequences, which indicates that the quality of the isolated DNA is adequate for amplification. The nptⅡ specific band has been amplified in three soil samples from the transgenic trees and even treated with filtration before the DNA isolation. This indicates that nptII DNA exists in the soil, although it is still unclear whether the DNA was in the soil particles, in the soil bacteria or in the Agrobacterium comamination which was used for the E. camaldulensis transformation. Two approaches on isolation of bacterial DNA have been suggested for testing the possibility of this event in the future.
