Fouts D E, Celander D W
Department of Microbiology, University of Illinois, Urbana 61801, USA.
Nucleic Acids Res. 1996 Apr 15;24(8):1582-4. doi: 10.1093/nar/24.8.1582.
RNA challenge phages are modified versions of bacteriophage P22 that allow one to select directly for a specific RNA-protein interaction in vivo. The original construction method for generating a bacteriophage that encodes a specific RNA target requires two homologous recombination reactions between plasmids and phages in bacteria. An improved method is described that enables one to readily construct RNA challenge phages through a single homologous recombination reaction in vivo. We have applied the new method to construct a derivative of P22R17, an RNA challenge phage that undergoes lysogenic development in bacterial cells that express the bacteriophage R17/MS2 coat protein.
RNA 挑战噬菌体是噬菌体 P22 的修饰版本，可用于在体内直接选择特定的 RNA-蛋白质相互作用。生成编码特定 RNA 靶标的噬菌体的原始构建方法需要在细菌中质粒与噬菌体之间进行两次同源重组反应。本文描述了一种改进方法，该方法能够通过体内单次同源重组反应轻松构建 RNA 挑战噬菌体。我们已应用该新方法构建了 P22R17 的衍生物，P22R17 是一种 RNA 挑战噬菌体，在表达噬菌体 R17/MS2 外壳蛋白的细菌细胞中经历溶原性发育。
