A DNA part library for reliable engineering of the emerging model nematode symbiotic bacteriumXenorhabdus griffiniaeHGB2511

Xenorhabdus griffiniaeis a bacterium that lives inside the intestine of the entomopathogenic nematodeSteinernema hermaphroditumand partners with the nematode to infect and kill insect larvae in soil. The construction of gene circuits, like reporters, inX. griffiniaewould provide tools to study and better understand the symbiotic relationship it has with its host. However, becauseX. griffiniaeis not a model organism, information about gene circuit construction inX. griffiniaeis limited. We develop and characterize a DNA part library similar to the CIDAR MoClo extension library forE. colito allow more efficient construction of genetic circuits inX. griffiniae. TurboRFP expressing strains with different constitutive Anderson promoters and different ribosome binding sites (RBS) were constructed to quantify promoter and RBS strengths inX. griffiniae. Furthermore, two fluorescent proteins sfGFP and sfYFP, as well as the bioluminescentluxCDABEoperon were added to the part library and successfully expressed inX. griffiniae. We then used the characterized parts to build and characterize IPTG inducible constructs.