一种改造载体多克隆位点的新方法

A Novel Method of the Reconstruction of Vector Multiple Cloning Sites

目的:探寻一种快速的、便捷的改造已知载体多克隆位点的方法。方法:应用一对5’末端分别添加了多个酶切位点的引物扩增任一已知DNA片段,用两端的限制性酶酶切后连接至载体多克隆位点中,从而引入两个新的酶切位点。我们将这一方法称为衔接子引物-PCR引入法。结果:两个研究实例的酶切结果证实:结果预先设计的酶切位点已替换至载体的多克隆位点,且引入的酶切位点能被顺利切割。结论:衔接子引物-PCR引入法是一种改造已知载体多克隆位点的可行方法。

Objective：Finding a way to reconstruct the vector MCS rapidly and easily. Method：The DNA fragment was amplified with prim- er pairs, whose 5＇ ends with several restriction sites. Then the DNA fragment was inserted into MCS of vector after digestion with endonu- clease in its both sides. This method was named adaptor primer -PCR introducing. Result：Digestion with endonuclease prove that the de- signed restriction sites have been inserted into MCS of the vector and the restriction sites were digestible, by using two examples. Conclusion：The adaptor primer - PCR introducing method was a feasible technique to reconstruct the vector MCS.