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目的 为了快速克隆ＰＣＲ产物，本实验构建了简单、方便的 ＴＡ载体。方法 ＰＢｌｕｅｓｃｒｉｐｔ ＳＫ（－）经 ＥｃｏＲ Ｖ酶切形成平端切口，在ＴａｑＥ的催化下加一个ｄＴＭＰ形成３’端带有Ｔ的粘端载体，在Ｔ４ＤＮＡ连接酶作用下，将ＰＣＲ产物互补连接于ＴＡ载体上，重组质粒转化细菌，碱裂解法提取重组质粒ＤＮＡ，酶切及电泳鉴定。结果ＴＡ载体成功地克隆了ＰＣＲ产物。结论构建的ＴＡ载体简单、方便，可以克隆任何ＰＣＲ产物。
Objective In order to rapidly clone PCR products, this experiment constructed a simple and convenient TA vector. Methods PBluescript SK (-) was digested with EcoR V to form a blunt nick end, and a dTMP was added under the catalysis of TaqE to form a 3 ’T vector with T terminus. The PCR product was ligated to the TA vector under the action of T4 DNA ligase The recombinant plasmids were transformed into bacteria, alkaline lysis method to extract the recombinant plasmid DNA, digested and electrophoresed. Results The TA vector successfully cloned the PCR product. Conclusion The constructed TA vector is simple and convenient, and any PCR product can be cloned.