| This paper characterized the gene of Ascorbate peroxidase(APXa) from rice under carbonate stress,which was made by carbonate (NaHCO3 ,Na2CO3 ) on plant.1.According to the sequences released on the Genebank, we designed a pair of primers for cytoplasm ascorbate peroxidase (APXa) , which reduced the toxic hazard made by reactive oxygen species(ROS), and amplified its full length cDNA from the total RNA in Rice (JIYUJING 1) by RT-PCR. Then APXa was linked into vector pMD18-T, after sequencing, It was linked not only midi-vector pTRIPLEX2 with multiple-cloning sites, but also downstream to Camv35S promoter and upstream to NOS terminator in PBI121, which was binary expression vector PBI121-APXa for plant, PCR and restriction endonuclease digestion were followed to detect the target. The recombined plasmid PBI121-APXa was transferred into Agrobacterium tumefacien EHA105 by electroshock.2.After a period of culturing and selecting, some transgenic plantlets resistant to kanamycin were obtained by Agrobacterium-mediated transformation from leaf discs under the kanamycin pressure, and four types of plantlets showed positive validated by detection of PCR and Southern .3.It proved that the APXa gene was integrated into the genome of tobacco.4.After being cultured in green-house for 3 months , the transformants showed sterility, for the pistil was higher than anther. We assumed that such variation was made by in vitro gene being integrated into the genome of tobacco... |
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