几种植物转基因表达载体的构建方法

Approaches of Constructing Expressional Vectors Utilized in Plant Genetic Transformation

表达载体的构建在植物基因工程研究中占据重要地位,直接关系到目的基因的表达效果。作者在收集整理前人的工作基础上,比较分析了5种不同载体构建方法的特点,给出了不同载体构建方法的适用性建议,期望能对植物基因工程中的载体构建工作提供有益帮助。在构建多片段连接的小载体的时候推荐用一步克隆法,在构建多片段连接的复杂的大载体时采用相应的复杂载体构建技术,在已获得无选择标记的转基因植株时采用三段T-DNA构建方法构建载体。在做基因功能验证时采用的Gateway技术,非常简单的载体构建可以采用传统的酶切连接的构建方式。现在的主流构建载体方式是利用结合其他手段的Gateway技术,未来载体的发展趋势将是无酶连接。

Construction of expression vector plays an important role in plant gene engineering, and is closely related with the expressional results of target geue. Based on the previous research work, the five different kinds of vector construction methods were summarized depending on their construction strategy. The suggestions have been proposed, aiming at specific plant expressional vector, to utilize the different construction strategy＇. Construction of multi - link in a serials of small fragment of vector cloning method recommended by one step methods, when building a complex multi - link fragments of the large vector using the complex constructing vector technology, in order to get the marker - free transgenic plants it can be used three T - DNAs vector construction methods. Adoption of the Gateway technology in doing functional verification, constructing very. simple vector can be using the traditional method of construction though digestion and connection. The mainstream way of construction vectors now is to use Gateway technology combing with other means, the development trend of the future constructing vector will be on no enzyme connection. At the same time, the tendency of plant vector construction in the future was prospected. It is hoped to promote the understandings of plant expressional vector construction.