Project description:Three-day metatranscriptome of surface gravel plain soils from the Central Namib Desert. Samples were collected at four times (6:00, 12:00, 18:00 and 24:00h) on each day (n=12). rRNA-depleted RNA was used to construct stranded libraries with the ScriptSeq v2 complete kit (Epicentre) adding unique barcodes in TruSeq adapters (ScriptSeq Index PCR primers, set 1, Epicentre). Libraries were single-end sequenced in a NextSeq 500 v2 sequencer, with read length of 75bp.
Project description:Purpose: To study the differential expression of gut bacterial mRNAs during chronic organophosphate treatment. Methods: Balb/c mice were treated with organophosphate (monocrotophos 28 ug/kg body weight/day) directly in drinking water for 180 days. Glucose tolerance tests indicated the induction of glucose intolerance. The total RNA was isolated by using TRIZOL from the cecal tissue that is cleared off fecal contents and subsequently the eukaryotic and bacterial rRNAs were removed by using MicrobExpress and MicrobEnrich kits (Ambion). Subsequently, RNA library was contructed using illumina kit as per manufacturer's instructions. Results: The reads were annotated to the reference human gut microbiome database and we found differential expression of large number of genes especially those involved in organophosphate degradation. Our subsequent studies proved that gut microbial degradation of organophosphates induces glucose intolerance via gluconeogenesis. Conclusions: Chronic organophosphate-induced hyperglycemia is mediated by the organosphosphate-degrading potential of gut microbiota.
