Activation of three natural product biosynthetic gene clusters from CGMCC 4.1386 by a reporter-guided strategy.

Along with the fast developing of DNA sequencing technology, a great number of natural product biosynthetic gene clusters have been discovered by bioinformatic analysis, which demands novel high-throughput genome mining methods to obtain the diverse compounds dictated by those gene clusters. In this work, a method based on the reporter gene was established to screen for the activation conditions of thirteen different gene clusters from CGMCC 4.1386. In this reporter-guided method, the key structure gene was replaced by a - cassette with the gene being controlled by the transcription and translation machinery of the key structure gene. It not only facilitated the screening of activation conditions, but also provided the null mutants of specific natural product gene clusters as controls to link those clusters with their products conveniently. The potential activation conditions of eleven gene clusters from CGMCC 4.1386 were obtained. In addition, activation of three of the eleven gene clusters was confirmed and their products were identified.