Nano-screening of marine symbiotic consortium for useful bio-materials and their analyses

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Recently, marine microbial consortium has been paid attention as new targets for screening of biologically active agents and so on. However, most of bacteria in environment are unculturable and unidentified. In this study, we aimed to have gene resources and develop the micro-assay system for small number of cells which are directly collected. investigate biodiversity, species identification of bacteria associated with marine sponge and coral, which are well known to produce several biologically active agents. Furthermore, methods for their efficient collection from host cells and genome amplification, and micro-screening system have been developed.Microbial cells were sorted by using flow cytometer and their 16SrDNA analyses were performed before/after cell sorting. 16SrDNA analysis showed that various kinds of bacteria which were not found in the sample not sorted were observed in the sorted cell samples. Those cells are not enough number of cells for construction of genome library. They are subjected to genome amplification by MDA method using phai-29 DNA polymerase. It was confirmed that enough genome DNA was amplified and amplicons less than 10 Kb were not chimeric when the model microbes were used. Furthermore, amine-coated nano-beads showed high performance in DNA extraction/collection.In order to micro-screening of useful materials, several devises were constructed. Cell-capture devise constructed showed 90% cell capture without any non-specific adsorption. Furthermore, micro-capillary devise for micro-screening was also constructed. In the devise, gradient of chemicals introduced into capillary was observed. This showed that it is possible to examine biological effect of target chemicals under the various of their concentration.