Bacillus subtilis genome vector-based engineering of genomic DNA

Technological developments in chromosome engineering are essential for the manipulation and functional analysis of genomic DNA fragments. In this study, we demonstrated the ability of the Bacillus subtilis genome (BGM) vector as an alternative platform for engineering large DNA fragments to modify and reconstruct genomic structure and to generate transgenic mice. The BGM vector system can utilize valuable BAC resources by a single-step transfer of the BAC insert into the BGM vector. We showed that this system enables complete genetic modifications of cloned DNA fragments using various homologous recombination methods, including insertion, deletion, inversion and connection to elongate a DNA fragment, without leaving any traces of modifications. We verified the sequences of the original BAC clones and the modified BGM inserts.