Selection of RNA-binding peptides using mRNA-peptide fusions.
We have been working to apply in vitro selection to isolate novel RNA-binding peptides. To do this, we use mRNA-protein fusions, peptides covalently attached to their own mRNA. Here, we report selection protocols developed using the arginine-rich domain of bacteriophage lambda-N protein and its binding target, the boxB RNA. Systematic investigation of possible paths for a selection round has allowed us to design a reliable and efficient protocol to enrich RNA-binding peptides from nonfunctional members of a complex mixture. The protocols we have developed should greatly facilitate the isolation of new molecules using the fusion system.
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
J E
JE
Barrick
T T
TT
Takahashi
A
A
Balakin
R W
RW
Roberts
D018076:DNA, Complementary / Q000378:metabolism; D005821:Genetic Techniques; D010455:Peptides; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*
D018076:DNA, Complementary; D010455:Peptides; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins