An efficient system for stable markerless integration of large biosynthetic gene clusters into Streptomyces chromosomes.

An efficient system for stable markerless integration of large biosynthetic gene clusters into Streptomyces chromosomes.
Appl Microbiol Biotechnol. 2021 Feb 10;:
Authors: Csolleiova D, Knirschova R, Rezuchova B, Homerova D, Sevcikova B, Matulova M, Núñez LE, Novakova R, Feckova L, Javorova R, Cortés J, Kormanec J
Abstract
The bacteria of the genus Streptomyces are among the most important producers of biologically active secondary metabolites. Moreover, recent genomic sequence data have shown their enormous genetic potential for new natural products, although many new biosynthetic gene clusters (BGCs) are silent. Therefore, efficient and stable genome modification techniques are needed to activate their production or to manipulate their biosynthesis towards increased production or improved properties. We have recently developed an efficient markerless genome modification system for streptomycetes based on positive blue/white selection of double crossovers using the bpsA gene from indigoidine biosynthesis, which has been successfully applied for markerless deletions of genes and BGCs. In the present study, we optimized this system for markerless insertion of large BGCs. In a pilot test experiment, we successfully inserted a part of the landomycin BGC (lanFABCDL) under the control of the ermEp* promoter in place of the actinorhodin BGC (act) of Streptomyces lividans TK24 and RedStrep 1.3. The resulting strains correctly produced UWM...

Source: Applied Microbiology and Biotechnology - February 10, 2021 Category: Microbiology Authors: Csolleiova D, Knirschova R, Rezuchova B, Homerova D, Sevcikova B, Matulova M, Núñez LE, Novakova R, Feckova L, Javorova R, Cortés J, Kormanec J Tags: Appl Microbiol Biotechnol Source Type: research