Application of Recombinase-Based In Vivo Expression Technology to <i>Bifidobacterium longum</i> subsp. <i>longum</i> for Identification of Genes Induced in the Gastrointestinal Tract of Mice

Abstract

Bifidobacteria are one of the major components in human gut microbiota and well-known as beneficial microbes. However, clarification of commensal mechanisms of bifidobacteria in the intestines is still ongoing, especially in the presence of the gut microbiota. Here, we applied recombinase-based in vivo expression technology (R-IVET) using the bacteriophage P1 Cre/loxP system to Bifidobacterium longum subsp. longum 105-A (B. longum 105-A) to identify genes that are specifically expressed in the gastrointestinal tract of conventionally raised mice. Oral administration of the genomic DNA library of B. longum 105-A to conventionally raised mice resulted in the identification of 73 in vivo-induced genes. Four out of seven tested genes were verified in vivo-specific induction at least in the cecum by quantitative reverse transcription PCR. Although there is still room for improvement of the system, our findings can contribute to expanding our understanding of the commensal behavior of B. longum in the gut ecosystem.

Keywords

• bifidobacteria
• <i>bifidobacterium longum</i> subsp. <i>longum</i>
• r-ivet
• in vivo gene expression
• cre recombinase
• gut microbiota
• qrt-pcr