16s rrna gene-targeted pcr/pyrosequencing  (Pyrosequencing Inc)

Journal: eLife

Article Title: Linalool combats Saprolegnia parasitica infections through direct killing of microbes and modulation of host immune system

doi: 10.7554/eLife.100393

Figure Lengend Snippet: In vitro, (1) Linalool influenced DNA transcription, tRNA transport, rRNA processing and maturation (5.8S, 18S, and 25S), and the biogenesis and assembly of ribosome subunits (40S and 60S) in the cell, which might lead to the reduction of S. parasitica growth; (2) Linalool disrupted the cell membrane, and the upregulation of glycerophospholipid metabolism likely represents the cell’s response to cope with this damage; (3) ABC transporters contributed to metabolic resistance by pumping linalool out of the cell. In vivo, (1) Linalool enhanced the complement and coagulation system which in turn activated host immune defense and lysate S. parasitica cells; (2) Linalool promoted wound healing, tissue repair, and phagocytosis to cope with S. parasitica infection; (3) Linalool positively modulated the immune response by increasing the abundance of beneficial Actinobacteriota; (4) Linalool stimulated the production of inflammatory cytokines ( il-1β and il-6 ) and chemokines ( ccl19 and ccl5 ) to lyse S. parasitica cells.

Article Snippet: Genomic DNA was extracted from grass carp intestinal content samples and stored at –20°C following the instructions of the 16S rRNA gene pyrosequencing DNA kit.

Techniques: In Vitro, Membrane, In Vivo, Coagulation, Infection