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.
Barrick
J E
JE
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Takahashi
T T
TT
Balakin
A
A
Roberts
R W
RW
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