一种共沉淀回收构建载体的新方法

A Co-precipitation Recover New Method for Constructing the Vectors

[目的]介绍一种新型的载体构建方法。[方法]利用乙醇可以使酶切反应中内切酶变性失活的特性对酶切产物进行回收和纯化，再将得到的混合沉淀溶解进行连接、转化。[结果]16—20h长出单菌落进行PCR鉴定，转化后的单菌落有10％的阳性率，将阳性菌落提取质粒进行酶切鉴定，电泳条带正确，载体构建成功。[结论]利用产物混合回收方法成功构建了植物表达载体pCambia1301cry3Aa，且省去酶切步骤，证明该方法是一种简便、实用的方法，具有一定的经济价值。

[ Objective ] The study described a novel method of constructing the vector. [ Method] This method was traditionally obtained after restriction enzyme digestion products and carrier are mixed precipitation recovery,,connection, transformation heat shock method [ Result ] Single colonies growing 16 -20 h were identified by PCR, and a single colony has transformed the positive rate of 10%. The positive colonies were men- tioning plasmid restriction enzyme digestion, and electrophoresis bands correct vector was successfully constructed. [ Conclusion ] Mixed use prod- uct recovery method successfully constructed plant expression vector pCambial3Ol-cry3Aa, and eliminating the need for digestion step. The study proved that this was a simple,practical method, and there was a certain economic value.