一种快速构建35S组成型植物双价表达载体的方法

A Fast Method of Constructing 35S Constitutive Plant Dual-gene Expresses Vectors

通过对几种常用植物表达载体的35S和Tnos序列进行比对,设计带有酶切位点的引物35S和Tnos,通过PCR克隆了同时带有35S和Tnos序列的目的基因片段,通过定向的酶切、连接获得双价表达载体。此方法简化了植物双价表达载体构建过程,具有明显时间短、效率高、不需要中间载体、检测简单等特点,而且引物对于多数常用35S组成型植物表达载体是通用的,为快速构建35S组成型植物双价表达载体提供了一种新方法。

This article reported a simple,general,effective dual-gene expression vector construction method. After comparing 35S and Trios sequence with several kinds of commonly plants expresses vectors,35S and Trios primers with restriction enzyme cutting sites were designed. Then the gene fragment with aim gene as well as 35S and the Tnos were cloned by using PCR and the directional enzyme cutting and connection were used to acquire the dual-gene expresses vectors. This method which simplified the process of plant dual-gene expresses vectors construction,has apparent improved qualifies,such as short period,high efficiency,no intermediate vectors needed,and easy-testing.