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Metal‐ion‐induced expression of gene fragments from subseafloor micro‐organisms in the Kumano forearc basin, Nankai Trough
Author(s) -
Wakamatsu T.,
Morono Y.,
Futagami T.,
Terada T.,
Nishikawa S.,
Morisawa T.,
Ohshita K.,
Inagaki F.,
Ashiuchi M.
Publication year - 2018
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.14061
Subject(s) - gene , gene expression , biology , microbiology and biotechnology , cell sorting , genetics , flow cytometry
Aims Using substrate‐induced gene‐expression ( SIGEX ) screening on subseafloor sediment samples from the Nankai Trough, Japan, we identified gene fragments showing an induction response to metal ions. Methods and Results Environmental DNA libraries in Escherichia coli host cells were tested by the addition of metal ions (Ni 2+ , Co 2+ , Ga 3+ or Mo 6+ ), followed by cell sorting of clones exhibiting green fluorescence upon co‐expression of green fluorescence protein downstream of the inserted gene fragments. One clone displayed Ni 2+ ‐specific induction, three clones displayed Ga 3+ ‐specific induction and three clones displayed an induction response to multiple metal ions. DNA sequence analysis showed that a variety of genes showed induction responses in the screened clones. Conclusions Using the SIGEX approach, we retrieved gene fragments with no previously identified response to metal ions that exhibited metal‐ion‐induced expression. This method has the potential to promote exploration of gene function through gene‐induction response. Significance and Impact of the Study We successfully linked gene‐induction response with sequence information for gene fragments of previously unknown function. The SIGEX ‐based approach exhibited the potential to identify genetic function in unknown gene pools from the deep subseafloor biosphere, as well as novel genetic components for future biotechnological applications.