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贾宇臣. 花生子叶节外植体遗传转化体系的建立[J]. 科学技术与工程, 2011, (14): .
jiayuchen.Establishment of Genetic Transformation System of Peanut Cotyledon Node Explants[J].Science Technology and Engineering,2011,(14):.
花生子叶节外植体遗传转化体系的建立
Establishment of Genetic Transformation System of Peanut Cotyledon Node Explants
投稿时间:2011-02-16  修订日期:2011-02-27
DOI:
中文关键词:  花生  遗传转化  农杆菌介导
英文关键词:peanut  Genetic transformation  Agrobacterium tumefaciens-mediated
基金项目:
  
作者单位
贾宇臣 内蒙古医学院
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中文摘要:
      目的: 为了建立花生的高效遗传转化体系。方法:利用花生子叶节高效再生体系,以农杆菌介导法,用含有表达质粒pBOG3VP7的根癌农杆菌进行外源基因转化,优化花生的遗传转化体系。结果:通过对影响农杆菌介导的花生遗传转化的诸多因素进行的优化,试验表明:侵染菌液添加100祄ol/L AS,结合负压处理,农杆菌侵染10min,共培养3d,延后选择2w。对抗性植株进行PCR和PCR-Southern检测,11株中有6株PCR反应呈阳性,其中有3株Southern杂交有特异性目标带出现。结论:结果表明,外源基因已经整合到了花生基因组上。
英文摘要:
      objective: In order to establish efficient genetic transformation system of peanuts. Method: Applying to regeneration system of peanut cotyledon node explants, the colony of LBA4404 containing pBOG3VP7 was used for thransformation. Genetic transformation system of peanut cotyledon nodes via Agrobacterium tumefaciens- mediated was developed, in which the effects of several factors influencing the tranformation were investigated and optimized. Result: Peanut cotyledon nodes were infected under the treatment of negative pressure for 10 min, co-cultured for 3 days, and regenerative shoots were screened on selective medium after sub-culture of 2 weeks. Concentration of AS with 100祄ol/L was helpful for successful infection. 6 plants among 11 plants which can grow up through selection on medium containing 125mgl-1 Kanamycin were assessed by PCR amplification .Then the genomic DNA of PCR positive plants were further analyzed with PCR-Southern blot hybridizations. 3 plants had all DNA blotting band. Conclusion: The results showed that the foreign gene has been integrated into genome of transformated peanuts.
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