Project description:Bacteriophages (phages) are widespread in Streptococcus pneumoniae, with most strains carrying phage genomes integrated into the chromosome. RNA sequencing was utilised to explore whether phage gene expression could be detected. The pneumococcal reference strain PMEN3 (Spain9V-3), which contained two full-length phages and one partial phage, was grown in broth culture and mitomycin C was added to facilitate phage induction. PMEN3 culture samples were taken at sequential time points and RNA was extracted and sequenced.
Project description:In this research, we used RNA-sequencing technology to detect genome-wide differentially expressed genes in spleen and gill of Vibrio harveyi -infected Takifugu rubripes.This high-throughput sequencing could help us to understand new mechanisms of action of V. harveyi induced aquaculture fish disease.
Project description:Vibrio harveyi, a significant pathogen in aquaculture, can lead to severe gut damage and metabolic disorders in fish. Verification via Transcriptome Sequencing of the gut damage caused by Vibrio harveyi in Hybrid Groupers and the Alleviating Effect of oligochitosan Feeding.
Project description:Vibrio harveyi, a significant pathogen in aquaculture, can lead to severe liver damage and metabolic disorders in fish. Verification via Transcriptome Sequencing of the Hepatic Damage Caused by Vibrio harveyi in Hybrid Groupers and the Alleviating Effect of Chitosan Oligosaccharide Feeding.
Project description:Whole-genome sequencing is an important way to understand the genetic information, gene function, biological characteristics, and living mechanisms of organisms. There is no difficulty to have mega-level genomes sequenced at present. However, we encountered a hard-to-sequence genome of Pseudomonas aeruginosa phage PaP1. The shotgun sequencing method failed to dissect this genome. After insisting for 10 years and going over 3 generations of sequencing techniques, we successfully dissected the PaP1 genome with 91,715 bp in length. Single-molecule sequencing revealed that this genome contains lots of modified bases, including 51 N6-methyladenines (m6A) and 152 N4-methylcytosines (m4C). At the same time, further investigations revealed a novel immune mechanism of bacteria, by which the host bacteria can recognize and repel the modified bases containing inserts in large scale, and this led to the failure of the shotgun method in PaP1 genome sequencing. Strategy of resolving this problem is use of non-library dependent sequencing techniques or use of the nfi- mutant of E. coli DH5M-NM-1 as the host bacteria to construct the shotgun library. In conclusion, we unlock the mystery of phage PaP1 genome hard to be sequenced, and discover a new mechanism of bacterial immunity in present study. Methylation profiling of Pseudomonas aeruginosa phage PaP1 using kinetic data generated by single-molecule, real-time (SMRT) sequencing on the PacBio RS.