Identification of novel broad host-range promoter sequences functional in diversePseudomonadotaby a promoter-trap approach

Abstract

AbstractFinding novel promoter sequences is a cornerstone of synthetic biology. To contribute to the expanding catalog of biological parts, we employed a promoter-trap approach to identify novel sequences within an Antarctic microbial community that act as broad host-range promoters functional in diversePseudomonadota. UsingPseudomonas putidaKT2440 as host, we generated a library comprising approximately 2,000 clones resulting in the identification of thirteen functional promoter sequences, thereby expanding the genetic toolkit available for this chassis. Some of the discovered promoter sequences prove to be broad host-range as they drove gene expression not only inP. putidaKT2440 but also inEscherichia coliDH5α,Cupriavidus taiwanensisR1T,Paraburkholderia phymatumSTM 815T,Ensifer meliloti1021, and an indigenous Antarctic bacterium,Pseudomonassp. UYIF39. Our findings enrich the existing catalog of biological parts, offering a repertoire of broad host-range promoter sequences that exhibit functionality across diverse members of the phylumPseudomonadota,proving Antarctic microbial community as a valuable resource for prospecting new biological parts for synthetic biology.