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将马铃薯X病毒(potato virus X,PVX)外壳蛋白(coat protein,CP)基因(cp)编码序列中植物偏爱密码子替换成稀有同义密码子,并将此修饰的外壳蛋白基因(cpm)及未修饰外壳蛋白基因(cp)分别置于CaMV 35S启动子后,构建植物表达载体,并用农杆菌介导转化烟草。Northern杂交及Run on实验结果表明,转cpm基因烟草比转cp基因烟草发生转录后基因沉默(PTGS)的频率明显增高(从6.25％增至35％),并且对人工接种的PVX病毒抗性显著提高。以上结果表明,基因中密码子的替换可以明显提高转基因植株的病毒抗性。
The coding sequence of plant protein gene (cp) in potato virus X (PVX) was replaced by rare codons, and the modified coat protein gene (cpm) and The unmodified coat protein gene (cp) was placed in the CaMV 35S promoter respectively to construct plant expression vector and Agrobacterium-mediated transformation of tobacco. Northern hybridization and Run on experiments showed that the frequency of posttranscriptional gene silencing (PTGS) was significantly increased (from 6.25% to 35%) in cpm transgenic tobacco than in cp transgenic tobacco, and was significantly resistant to PVX virus inoculation improve. The above results show that codon replacement in the gene can significantly increase the virus resistance of the transgenic plants.