A DNA Part Library for Reliable Engineering of the Emerging Model Nematode Symbiotic Bacterium <i>Xenorhabdus griffiniae</i> HGB2511

<i>Xenorhabdus griffiniae</i> is a bacterium that lives inside the intestine of the entomopathogenic nematode <i>Steinernema hermaphroditum</i> and partners with the nematode to infect and kill insect larvae in soil. The construction of gene circuits, such as reporters, in <i>X. griffiniae</i> would provide tools to study and better understand the symbiotic relationship it has with its host. However, because <i>X. griffiniae</i> is not a model organism, information about gene circuit construction in <i>X. griffiniae</i> is limited. We developed and characterized a DNA part library similar to the CIDAR MoClo extension library for <i>E. coli</i> to allow more efficient construction of genetic circuits in <i>X. griffiniae</i>. TurboRFP expressing strains with different constitutive Anderson promoters and different ribosome binding sites (RBS) were constructed to quantify promoter and RBS strengths in <i>X. griffiniae</i>. Furthermore, two fluorescent proteins sfGFP and sfYFP as well as the bioluminescent <i>luxCDABE</i> operon were added to the part library and successfully expressed in <i>X. griffiniae</i>. We then used the characterized parts of the cell to build and characterize IPTG inducible constructs.