[BBG02-P02] Screening of metal-ion inducible genes from subseafloor sedeiments of Nankai Trouph using substrate-induced gene expression method
Keywords:Deap-sea sediment, Gene function, Nankai Trough, Metagenome, SIGEX, Metal-ion
Constructed metagenomics shotgun libraries that confers gfp gene downstream of the inserted DNA fragment were induced by metal ions including Ni2+, Co2+, Mg2+, Mn2+, Mo6+, and Ga3+, followed by sorting of clones with green fluorescence by a cell sorter. From sorted clones, we secondary screened clones for its response to the induction of each metal ion. Clones isolated by using Ni2+, Ga3+ showed specific response to each metal ion used for isolation. Also there were other type of clones that showed response to induction by multiple metal ions. DNA sequence analysis revealed that while around half of the clones had database-identifiable DNA fragment, other half did not show any match to DNA sequences in database. This study showed great potential of SIGEX-based approach to find function of DNA fragments from large pool on unknown environmental DNA of natural environment.