This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The linear expression of a given gene can be interrupted at two levels (RNA and protein) by unique segments of information encoded by the gene. Here we report on a novel splicing process of an engineered protein in Escherichia coli at the entry to stationary phase of growth. This process differs from the known protein splicing mechanism acting through inteins as follows: (a) the internal excised domain is a peptide, 21 amino acids long, that we call an """"""""intide"""""""" (internal peptide);b) the nature of the amino acids inside the intide is not important, while the size of the intide is;(c) specific amino acids, i.e., RG upstream and KA downstream from the intide, have to be located 3 to 4 amino acids away from the borders of the intide;and (d) the process is growth-phase dependent.
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