Project description:To investigate the effect of CodY mutation on the gene expression in Streptococcus suis serotype 2 SC19 strain, we have employed whole genome microarray expression profiling as a discovery platform to identify genes regulated by CodY mutation. DNA microarray analysis was performed using an Agilent custom-designed oligonucleotide microarray. Based upon the whole genome sequence of SC19 , specific 60-mer oligonucleotide probes were designed using eArray (https://earray.chem.agilent.com/earray/), to cover all annotated genes. Probes were printed seven times on microarray slides. Three biological replicates of total RNA from two wild type strains and from two codY mutant strains were amplified and labeled with Cy3-CTP using Low Input Quick Amp Labeling Kit, one-color(Agilent technologies, US), following the manufacturer’s instructions. Labeled cRNA was purified using the RNeasy mini kit (Qiagen). After fragmentation, microarray slides were hybridized with 600 ng Cy3-labeled cRNA. Hybridization was performed at 65 °C for 17 h with rotation at 10 rpm. Microarray slides were washed and scanned by an Agilent Microarray Scanner (G2565BA). Those genes with greater than two-fold change ratios were regarded as differentially expressed genes. codY mutation induced gene expression in Streptococcus suis serotype 2 SC19 was detected in two wild type and two codY mutated strain of Streptococcus suis serotype 2.
Project description:To investigate the effect of CodY mutation on the gene expression in Streptococcus suis serotype 2 SC19 strain, we have employed whole genome microarray expression profiling as a discovery platform to identify genes regulated by CodY mutation. DNA microarray analysis was performed using an Agilent custom-designed oligonucleotide microarray. Based upon the whole genome sequence of SC19 , specific 60-mer oligonucleotide probes were designed using eArray (https://earray.chem.agilent.com/earray/), to cover all annotated genes. Probes were printed seven times on microarray slides. Three biological replicates of total RNA from two wild type strains and from two codY mutant strains were amplified and labeled with Cy3-CTP using Low Input Quick Amp Labeling Kit, one-color(Agilent technologies, US), following the manufacturer’s instructions. Labeled cRNA was purified using the RNeasy mini kit (Qiagen). After fragmentation, microarray slides were hybridized with 600 ng Cy3-labeled cRNA. Hybridization was performed at 65 °C for 17 h with rotation at 10 rpm. Microarray slides were washed and scanned by an Agilent Microarray Scanner (G2565BA). Those genes with greater than two-fold change ratios were regarded as differentially expressed genes.
Project description:Streptococcus suis is an important zoonosis pathogen that causes significant economic losses worldwide characterized by meningitis, septicaemia, arthritis, bronchopneumonia endocarditis. Streptcoccus suis 2 strain SC19 was isolated in Sichuan province in China, during the outbreak in 2005. Septicemia is most popular symptoms for SC19 infection, and mortality is high. We used human acute monocytic leukemia cell line (THP-1) infected SC19 to analysis the pathomechanism of septicemia in SS2 infection.
Project description:Streptococcus suis serotype 2 is an important pathogen of pigs, and the disease it causes is characterized by meningitis, septicaemia and pneumonia with high mortality. The pathogen is also an emerging zoonotic agent and threatens humans that are exposed to pigs or their by-products. We investigated the response of PBMC (Peripheral Blood Mononuclear Cell), brain and lung tissues to infection with S. suis 2 strain SC19 by using the Affymetrix Porcine Genome Array.
Project description:Streptococcus suis serotype 2 is an important pathogen of pigs, and the disease it causes is characterized by meningitis, septicaemia and pneumonia with high mortality. The pathogen is also an emerging zoonotic agent and threatens humans that are exposed to pigs or their by-products. We investigated the response of PBMC (Peripheral Blood Mononuclear Cell), brain and lung tissues to infection with S. suis 2 strain SC19 by using the Affymetrix Porcine Genome Array. Six piglets free of S. suis 2 were allocated randomly to the infected group and the uninfected group. Each piglet of the infected group was intravenous injection with Streptococcus suis 2 strain SC19 at a dose of 3Ã105 colony-forming units (CFU). Each piglet of the noninfected group was treated similarly with an identical volume of PBS as control. At 24 h after challenge, the pigs were slaughtered and their brains, lungs and PBMC were collected with RNase-free equipment for microarray analysis.
Project description:Streptococcus suis is an important zoonosis pathogen that causes significant economic losses worldwide characterized by meningitis, septicaemia, arthritis, bronchopneumonia endocarditis. Streptcoccus suis 2 strain SC19 was isolated in Sichuan province in China, during the outbreak in 2005. Septicemia is most popular symptoms for SC19 infection, and mortality is high. We used human acute monocytic leukemia cell line (THP-1) infected SC19 to analysis the pathomechanism of septicemia in SS2 infection. Human acute monocytic leukemia cell line (THP-1) cells were stimulated with Streptcoccus suis 2 (SS2) strain SC19. We added SS2 to THP-1 cells at a MOI of 1:1 (bacteria/cells). Uninfected control cells were incubated with PBS only. After 3 hours incubation, cells were collected for RNA extraction and hybridization on Affymetrix microarrays. A total of 4 samples were challenged, and 4 samples were used as controls. 4 microarrays were used in this experiment.