Construction of RNAi vector which resists cucumber mosaic virus and transformation of tobacco | 抗黄瓜花叶病毒RNAi载体的构建及烟草的转化
2010
Zhang Yu, Shihezi University, Shihezi (China), College of Life Sciences | Zheng Yinying, Shihezi University, Shihezi (China), College of Life Sciences | Zhao Fengyu, Shihezi University, Shihezi (China), College of Life Sciences
[Objective] Aimed to construct RNAi vector of cucumber mosaic virus and transferred this vector into tobacco. [Method] RT-PCR method was used to amplify cucumber mosaic virus NS04 and process RNA2 gene sequen of tomato isolates. The analysis results of phylogenetic tree demonstrated that the sequence in RNA2 encoded CMV-2a had 98.0% and 96.5% homology with nucleotide and amino acid of DQ412731 isolate of Zhejiang, China. The replicase fragment in CMV RAN2 gene was taken as target sequence to construct pBi35SCR2 eukaryotic expression vector, then the expression vector was identified. Through agrobacterium-mediated method, the expression vector was transferred into tabacco and PCR method was used to check the transfer. The PCR results demonstrated that the experiment had successfully construct eukaryotic expression vector of pBi35SCR2 and the expression vector was successfully transferred into tabacco. [Conclusion] The obtained transgenic tobacco could be used as challenge test material in following experiment and provided foundation for studying processing tomato resist cucumber mosaic virus.
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