Studies on the Fidelity of Viral Protein Synthesis

Abstract

A method for peptide mapping of tryptic digests on cellulose this layers was developed. The method was much more sensitive than procedures based on paper or column separation techniques, and enabled the detection of many peptides resulting from non-specific tryptic hydrolysis during protein digestion. An E-coli cell-free protein synthesizing system directed by R17 RNA was used to examine the fidelity of translation in vitro. The major product of this system, the coat protein, was separated from the in vitro reaction mixtures and analysed by the peptide mapping procedure. A series of experiments utilising radioactive amino acids established that the fidelity of translation was high, and enabled the identification of the mapping position of the major tryptic peptides.