TA载体pEASY-T1 Simple在基因克隆中的新应用

New Application of TA Vector pEASY-T1 Simple in Gene Cloning

在基因克隆操作中,由于常用克隆载体多克隆位点的酶切位点数目有限,造成一些DNA片段的克隆、连接受限。为解决这一问题,本文利用pEASY-T1 S imp le载体无酶切位点的特点,通过TA克隆的方法引入多个单一酶切位点,并对所构建的克隆载体进行了验证。最后证明,这种方法可高效构建含有不同多克隆位点的载体,克服了对常用克隆载体的依赖,大大提高了基因操作的灵活性和有效性。

A major challenge in the cloning of multiple DNA fragments is to find suitable restriction enzyme sites in the commonly used cloning vectors. To address this problem, in this study, it is constructed a new TA cloning vectors by adding desirable multiple cloning sites （MCS） to the pEASY -T1 Simple, which do not contain any restriction enzyme sites in original forms. Subsequently, it is verified the new cloning vectors by digesting the designed MCS. The results show that the designed MCS were successfully engi- neered into the pEASY - T1 Simple vector. In summary, an efficient method has been developed to construel the personalized TA cloning vectors with desirable MCSs, and thus to improve the flexibility and efficiency of gene engineering.