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SRX13613936: GSM5772487: Macrophage-sopF-R1; Gallus gallus; RNA-Seq
1 ILLUMINA (NextSeq 500) run: 69.2M spots, 20.9G bases, 6.7Gb downloads

External Id: GSM5772487_r1
Submitted by: Discovery Bacteriology & Microbiome, Elanco Animal Health
Study: Transcriptomic analysis of attenuated Salmonella Typhimurium infected chicken primary macrophages
show Abstracthide Abstract
Salmonella enterica is one of the most important foodborne pathogens that infect a variety of animals and birds. In humans, S. Typhimurium causes gastroenteritis, leading to vomiting, diarrhea, fever, and abdominal cramps. We mainly get infected with Salmonella by ingesting comminated poultry products. Therefore, developing an oral live attenuated vaccine for the poultry industry is our best bet against Salmonella infection. In this article, we investigated the potential of the next generation of Salmonella vaccines. We generated a library of potentially attenuated S. Typhimurium mutants and compared fitness to that of a commercial vaccine. We also investigated the invasion and survival potential of these mutants in chicken macrophages. Our data indicate that although these mutants had no significant growth defects, they were much sensitive to macrophage attack. Analyzing the transcriptome data from infected primary chicken macrophages, we concluded that these mutants elicit a robust immune response by activating several immunoregulatory pathways. Our data also indicates that by combining phoPQ deletion with an already existing cya-crp deletion in MeganVac1, a much stronger immune response can be generated. Overall design: Transcriptome profile of attenuated Salmonella infected primary macrophages
Sample: Macrophage-sopF-R1
SAMN24648232 • SRS11503047 • All experiments • All runs
Organism: Gallus gallus
Library:
Name: GSM5772487
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: The samples were processed by Cofactor Genomics as described previously (do Amaral et al., 2020). Briefly, total RNA was isolated using Qiagen RNAeasy Mini Kit (Qiagen) following the manufacturer's instructions. The isolated RNA was then treated with DNase I (Thermo Fisher Scientific) to remove potential DNA contamination. RNA was then precipitated using lithium chloride (Sigma). The concentration and quality of RNA were determined using nanophotometer. RNA samples with RNA integrity number (RIN) > 8.0, A260/A280 > 1.9 and A260/A230 > 2 were selected for mRNA library preparation and Next Generation Sequencing. Briefly, the total RNA was incubated with mRNA capture beads to remove contaminating ribosomal RNA from the sample using the Kapa Stranded mRNA-Seq kit (Kapa Biosystems) following manufacturer's instructions. The resulting poly(A)-captured mRNA was then fragmented. First-strand cDNA synthesis was performed using reverse transcriptase and random primers in the presence of Actinomycin D, followed by second-strand cDNA synthesis with DNA polymerase I and RNase H. Double-stranded cDNA was end-repaired and A-tailed for subsequent adaptor ligation. Indexed adaptors were ligated to the A-tailed cDNA. Enrichment by PCR was performed to generate the final cDNA sequencing library. Libraries were sequenced as paired-end 150 base reads on an Illumina NextSeq 500 following the manufacturer's protocols.
Runs: 1 run, 69.2M spots, 20.9G bases, 6.7Gb
Run# of Spots# of BasesSizePublished
SRR1744267469,223,29420.9G6.7Gb2022-05-25

ID:
18922573

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