ENGINEERED TRANSPOSON FOR FACILE CONSTRUCTION OF A RANDOM PROTEIN DOMAIN INSERTION LIBRARY

摘要

Methods for facile construction of a random domain insertion library (1) with optimal control of composition and length of inter-domain linker residues and (2) mediated by sticky-end ligation between host and guest DNA fragments. To develop such a method, we engineered a Mu transposon. The method exploits transposition of the engineered Mu transposon, which, upon removal, allows for sticky-end ligation between host and guest DNA fragments. We used a gene coding for xylanase from bacillus circulans (BCX) as a guest DNA sequence and the plasmid PUC19 containing lacZα as the target for insertion (i.e., a host DNA sequence). Results demonstrate that the method enables facile construction of a random domain insertion library with optimal control of composition and length of inter-domain linker residues.