Our knowledge of biological systems relies increasingly on the ability of quantifying and imaging intracellular signals and events in living subjects. The development of novel methods and advances in biotechnology have provided many basic tools that allow analyses of the complex biological systems in living cells. Since the discovery of protein splicing in 1990, the elucidation of the splicing mechanism and the identification of key amino acid residues involved in the dissection and ligation of the peptide bonds have facilitated the molecular engineering of inteins for different applications in protein chemistry. These include protein purification, protein ligation and peptide cyclization, construction of split reporter proteins, regulation of protein activity, and introduction of non-natural amino acids. In this chapter, we focus on the construction of split reporter proteins and their applications for detecting protein-protein interactions, identification of organelle-localized proteins, growing safer transgenic plants, and screening antimycobacterial agents.

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