基于XcmⅠ的T载体及其在合成生物学中的应用

An XcmⅠ-generated T vector and its applications in synthetic biology

摘要:

为了节约成本和提高实验效率，以商用pMD18-T载体为基础，构建获得了pFL-XS-T载体，其具有符合Biobrick标准的串联XbaⅠ-XcmⅠ-XcmⅠ-SpeⅠ克隆序列，通过对XcmⅠ的酶切处理即可以通过TA克隆方便地克隆PCR片段。克隆验证实验结果表明，经XcmⅠ处理的该载体可以与PCR片段进行TA连接，连接效率及阳性率均可以满足实验室要求并且可以得到Biobrick标准质粒。此外，该载体不仅可以与其余Biobrick标准元件进行串联，还可以作为功能DNA元件的筛选载体。

Abstract:

For more economical and efficient DNA clonging, pFL-XS-T, a Biobrick-T vector was constructed based on pMD18-T vector, carrying clonging regions of XbaⅠ-XcmⅠ-XcmⅠ-SpeⅠ. The results revealed that PCR products could be conveniently inserted into pFL-XS-T vevtor digested by XcmⅠby means of TA cloning. The positive frequency of recombination can meet the experimental requirements and all the plasmids obtained meet Biobrick standard. Moreover, the pFL-XS-T is compatible with other Biobrick parts, and serves as a vector for functional DNA fragments screening.