ABSTRACT
Global analysis o f protein functions and networks has become the focus o f considerable attention since the sequencing o f the human genome. The development o f proteomics has led to an increasing interest in cell-free translation systems because o f their rapidity and ease o f handling. We have developed the so-called in vitro virus ( I W ) as a more stable and efficient tool for evolutionary protein engineering. This system is applicable as an m RNA display technique to analyze protein functions and networks in proteomics. Accordingly, we developed a high-throughput I W system for analysis o f protein-protein interactions employing cell-free co-translation and selection. Here, we overview this system and discuss the advantages o f analyz ing protein-protein interactions using I W as a genotype-phenotype assignment molecule in combination with cell-free co-translation. Moreover, we discuss how we address false positives and negatives in this system, and the in silico mass data processing required to derive biologically significant interactions from the wealth o f raw data.
