Improved method for selecting RNA-binding activities in vivo.
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.
Fouts
D E
DE
Department of Microbiology, University of Illinois, Urbana 61801, USA.
Celander
D W
DW
D017100:Bacteriophage P22 / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D011995:Recombination, Genetic
D012367:RNA, Viral; D016601:RNA-Binding Proteins