Project description:Gene content comparison of control C. jejuni subsp. jejuni strain 11168 which colonizes and causes disease in C57BL/6 IL-10-/- mice versus C. jejuni strains D6844, D6845, D6846, D6847, D6848, D6849, D0121, D0835, D2586, D2600,33560 and NW in the C57BL/6 IL-10-/- mice. Keywords: DNA/DNA comparison
Project description:A number of seven proteins were selected during immunoscreening and further analyses. The proteins were in silico divided into overlapping 15-mer oligopeptides with an overlap of 11 residues. The microarrays were incubated with different antibodies to C. jejuni, Escherichia coli and Salmonella enterica. Each microarray was separated into three individual incubation chambers using ProPlate 3-Well modules. Within each incubation chamber, each peptide was spotted in triplicate with the controls spotted nine times each. The controls included human-IgG, rabbit-IgG, mouse-IgG and myelin basal protein (MBP). Each chamber was incubated independently using different polyclonal antibodies to C. jejuni, and for specificity testing, with an antibody to E. coli or S. enterica. Thus, samples 4_1, 4_2, 5_1 and 5_2 represent epitope mapping of three proteins with C. jejuni antibodies, while 6_1, 6_2, 7_1 and 7_2 represent the data after incubation with an E. coli antibody investigating unspecific interactions of the antibody to the potential linear epitopes from C. jejuni. Finally, for four different proteins from C. jejuni, the set two indicated by S2 was performed. Here, S2_6_1, S2_7_1, S2_7_2, S2_8_1 and S2_8_2 indicate epitope mapping after incubation with antibodies to C. jejuni, while the remaining samples were performed to test these latter 4 proteins for specificity by incubation with antibody to S. enterica.
Project description:Gene content comparison of control C. jejuni subsp. jejuni strain 11168 which colonizes and causes disease in C57BL/6 IL-10-/- mice versus C. jejuni strains D6844, D6845, D6846, D6847, D6848, D6849, D0121, D0835, D2586, D2600,33560 and NW in the C57BL/6 IL-10-/- mice. Keywords: DNA/DNA comparison Two genome comparison of disease strain versus non disease strain of C.j., 4 Biological replicates - 2 of which were dye swaps
Project description:Reference and type strains of well-known bacteria have been a cornerstone of microbiology research for decades. The sharing of well-characterized isolates among laboratories has run in parallel with research efforts and enhanced the reproducibility of experiments, leading to a wealth of knowledge about trait variation in different species and the underlying genetics. Campylobacter jejuni strain NCTC 11168, deposited at the National Collection of Type Cultures in 1977, has been adopted widely as a reference strain by researchers worldwide and was the first Campylobacter for which the complete genome was published (in 2000). In this study, we collected 23 C. jejuni NCTC 11168 reference isolates from laboratories across the UK and compared variation in simple laboratory phenotypes with genetic variation in sequenced genomes. Putatively identical isolates, identified previously to have aberrant phenotypes, varied by up to 281 SNPs (in 15 genes) compared to the most recent reference strain. Isolates also display considerable phenotype variation in motility, morphology, growth at 37 °C, invasion of chicken and human cell lines, and susceptibility to ampicillin. This study provides evidence of ongoing evolutionary change among C. jejuni isolates as they are cultured in different laboratories and highlights the need for careful consideration of genetic variation within laboratory reference strains. This article contains data hosted by Microreact.
Project description:We report the use of differential RNA-sequencing for the determination of the primary transcriptome of wildtype Campylobacter jejuni NCTC 11168. This allows for the genome-wide determination of transcription start sites.