title: Specific binding of a chemically synthesized prokaryotic ribosome recognition site. Prospect for molecular cloning and expression of eukaryotic genes.
abstract: An icosadeoxyribonucleotide containing the several features found in prokaryotic mRNA ribosome binding sites has been synthesized. This sequence can stimulate the binding of initiator fMet-tRNAf to the ribosome to form a stable 71 S initiation complex identical with those induced by natural messengers. The binding of this synthetic ribosome binding site is absolutely dependent upon initiation factor IF3, and the bound fMet-tRNAf is sensitive to puromycin indicating the formation of a functional initiation complex. A heptadecadeoxyribonucleotide, identical with the icosanucleotide but lacking the terminal A-T-G codon, can also stimulate the stable binding of fMet-tRNAf to the ribosome, suggesting that the selection of the proper A-U-G initiation codon by fMet-tRNAf is subsequent to and a result of the recognition and binding of the fMet-tRNAf . 30 S ribosome complex to the initiation site. The prospect of ligating a similar synthetic ribosome binding site in front of a eukaryotic gene for cloning in an appropriate prokaryotic vector to assure the expresion of the protein is discussed.
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D008715:Methionine; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D012343:RNA, Transfer; D008715:Methionine