Chromatic Bacteria v.2-A Himar1 Transposon-Based Delivery Vector to Extend the Host Range of a Toolbox to Fluorescently Tag Bacteria

Abstract

A recent publication described the construction and utility of a comprehensive “Chromatic Bacteria” toolbox containing a set of genetic tools that allows for fluorescently tagging a variety of Proteobacteria. In an effort to expand the range of bacteria taggable with the Chromatic Bacteria toolbox, a series of Himar1 transposon vectors was constructed to mediate insertion of fluorescent protein and antibiotic resistant genes. The Himar1 transposon was chosen as it is known to function in a wide range of bacterial species. To test the suitability of the new Himar1 Chromatic Bacteria plasmid derivatives, conjugations were attempted on recently isolated non-model organisms. Although we were unsuccessful in delivering the plasmids into Gram-positive bacterial isolates, we successfully modified previously recalcitrant isolates to the first set of the Chromatic Bacteria toolbox, such as Sphingomonas sp. Leaf357 and Acidovorax sp. Leaf84. This manuscript reports on the currently available plasmids and transposition success in different bacteria.

A self‐propagating, barcoded transposon system for the dynamic rewiring of genomic networks

“…In this context, our platform exhibits several features that facilitate its extensibility to diverse bacterial strains and applications. First, the himar1C9 mariner transposase that is used in our platform is compatible with a broad range of bacterial hosts (Goodman et al , 2009 ; Perry & Yost, 2014 ; Stocks et al , 2022 ). Second, the modular assembly pipeline that is used to construct the donor plasmids enables the transposons to be functionalized with diverse genetic cargos and accelerates prototyping for diverse microbial hosts (Liu et al , 2018 ).…”