Benhar I, Engelberg-Kulka H
Department of Molecular Biology, Hebrew University-Hadassah Medical School, Jerusalem, Israel.
Gene. 1991 Jul 15;103(1):79-82. doi: 10.1016/0378-1119(91)90394-q.
We describe a novel procedure for determining the amino acid (aa) sequence of the internal regions of proteins. This procedure has been implemented by directly determining the sequence of aa 65-75 of the product of the trpR gene of Escherichia coli, the trp repressor. This method is based on the insertion of the cleavage site of a specific protease (factor Xa) into the protein immediately before the region to be sequenced by Edman degradation. The simplicity of the procedure makes it appealing for studies of protein structure-function relationships, and of the expression of genetic information. The method is particularly useful when there is ambiguity concerning the co-linearity of the aa and nucleotide sequences.
