Abstract
The generation of an engineering strain with reduced genome relies on efficient methods for marker-free gene knockout. In this study, research on the efficiency of deletion of genome fragments with upp-based counterselective method in Bacillus subtilis was carried out. Using this method, the 3.053 Kb, 12.414 Kb and 34.148 Kb fragments of the skin element were marker-free deleted successfully, with the efficiency of 27%, 11.4% and 25%, respectively. Here we demonstrate that this method is an efficient genetic tool for large genome fragments marker-free deletion.
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