应用Gateway技术构建水稻OsDAD1基因的RNA干涉载体

Construction of RNAi Vectors for OsDAD1 Gene of Rice Using Gateway Technology

近年来,RNA干涉广泛应用于植物基因功能研究。Gateway技术是通过采用多重载体高效构建目标基因及任何DNA片段克隆和表达载体的研究平台,该技术以λ噬菌体的位点特异性重组体系为基础,两端具有重组位点的DNA片段或目标基因可以非常容易地被重组克隆到含同源重组位点的载体上。这种以重组为基础的克隆体系目前已被广泛应用于分子生物学中来分析基因的功能。本文应用Gateway技术体系替代传统载体构建方法,构建了水稻基因OsDAD1的RNA干涉载体,从而证明了Gateway技术的可行性。

In the last decade, RNA interference （RNAi） is an effective strategy to be widely used to identify the function of genes in plants. Gateway technology is a high throughput cloning and expression platform that allows rapid and highly efficient cloning of targeted gene or any DNA segment across multiple vectors. It is based on the site-specific recombination reaction mediated by phage λ. DNA fragments or targeted gene flanked by recombination sites can be transferred into vectors that contain compatible recombination sites. This approach to cloning has transformed molecular biology research, facilitating analysis of gene function while providing significant timesavings over other methods. In this paper, Gateway technology is used to replace previously conventional cloning vector construct and an RNAi vector containing OsDAD1 gene was constructed, and this technique is proved to be a practicable way to RNAi construct.