一种稳定、易于操作的噬菌体随机肽库载体的构建

Establishment of a stable and easily manipulated vector for macrophage random peptide library

目的：拟将噬菌粒ｐＣＯＭＢ３改建为更为稳定、高拷贝且易于操作的噬菌体随机肽库载体。方法：对常用噬菌体抗体库呈现载体ｐＣＯＭＢ３序列进行点矩阵作图分析（Ｄｏｔ－Ｐｌｏｔ分析），切除其中重复区，纠正其多代培养后自删除特性。原克隆位点６３ｂｐ的填充序列增加到１４３５ｂｐ以便于酶切鉴定和电泳回收。结果：构建了命名为ｐＴＭＢ２的噬菌粒载体。结论：ｐＴＭＢ２的噬菌粒载体更为稳定且易于操作。

Objective: To modify a combinational library display vector, pCOMB3 and to provide a stable, easily manipulated and highly copied vector for the display of a random peptide library. Methods: According to Dot Plot analysis of pCOMB3 nucleic acid sequences, the phagmid pCOMB3 was modified to eliminate the propensity of accumulated deletions in the region of cloning site. Furthermore, a 1375 bp DNA fragment from phage was cloned in the small 63 bp stuffer at the cloning site. Results: A 5129 vector named pTMB2 was produced. Conclusion: pTMB2 holds the appropriate characteristics for random peptide phage display with improved stability and ease of manipulation.