转抗草甘膦基因烟草T-DNA侧翼序列的扩增与分析Amplification and Analysis of T-DNA Flanking Sequence in Transgenic Glyphosate-resistant Tobacco
李同亮;王坤波;宋国立;刘方;黎绍惠;王春英;张香娣;王玉红;
摘要(Abstract):
[目的]研究抗草甘膦基因在烟草中的插入位点、T-DNA结构等整合情况。[方法]利用现有的3个转抗草甘膦基因的烟草材料,通过PCR Walking的方法扩增出携带抗草甘膦基因的T-DNA侧翼序列,并用DNAMAN、BLAST等生物信息学方法进行分析。[结果]对获得的PCR条带进行分析,结果表明这些侧翼序列均含载体骨架序列且结构各不相同,T-DNA和载体骨架序列都存在重组现象。[结论]研究在转基因植株中检测到了载体的骨架结构,说明转基因的同时有可能会造成非目的基因的转移,即转基因植物存在一定的风险,应对转基因植物转化载体和基因工程技术路线进一步改良。
关键词(KeyWords): 转基因烟草;T-DNA;整合;侧翼序列
基金项目(Foundation): 国家转基因生物新品种培育重大专项(2008ZX08005003)
作者(Author): 李同亮;王坤波;宋国立;刘方;黎绍惠;王春英;张香娣;王玉红;
Email:
DOI: 10.13989/j.cnki.0517-6611.2009.21.075
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