High-throughput identification and validation of in situ-expressed genes of Lactococcus lactis

摘要: Understanding the functional response of bacteria to their natural environment is one current challenges in microbiology. Over past decades several techniques have been developed study gene expression complex habitats. Most these methods, however, are laborious, and validation results under situ conditions cumbersome. Here we report improvement recombinase-based vivo technology (R-IVET) by implementation two additional reporter genes. The first an α-galactosidase (melA), which facilitates rapid identification vivo-induced Second, bacterial luciferase genes (luxAB) transcriptionally coupled resolvase gene, allows characterization system implemented validated industrially important lactic acid bacterium Lactococcus lactis. We demonstrate applicability advanced R-IVET lactococcal promoter elements that induced minimal medium compared commonly used rich laboratory M17. screening led 19 promoters predominantly control involved amino nucleotide metabolism transport functions. Furthermore, high-resolution transcription analysis enabled constituents specific molecules control. Rapid target exemplifies high-throughput potential extended system. can be applied other species, provided organism interest.