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Identification of genes differentially expressed in RAW264.7 cells infected by Salmonella typhimurium using PCR method
Authors:Kang Kyung Ho  Song Jung A  Shin Dong-Jun  Choy Hyon E  Hong Yeongjin
Institution:Genomic Research Center for Enteropathogenic Bacteria, and Department of Microbiology, Chonnam National University Medical School, Gwangju 501-746, Republic of Korea.
Abstract:Salmonella typhimurium, causing mouse typhoid, infects hosts such as macrophage cells, and proliferates in intracellular vacuoles causing infected cells to trigger numerous genes to respond against the infection. In this study, we tried to identify such genes in RAW264.7 cells by using the PCR screening method with degenerate primers. Fourteen genes were found to be differentially expressed after a 4 h infection in which the expression of 8 genes increased while expression of the others decreased. Most of the genes were involved in proinflammatory responses such as cytokines production and cell death. The mutation in msbB gene encoding the myristoyl transferase in lipid A of lipopolysaccharide (LPS) resulted in much lower toxicity to the inoculated animals. We compared the expression of the identified genes in wild-type and msbB-mutated S. typhimurium infections and found that Lyzs encoding lysozyme type M was differentially expressed. This gene is quite likely to be related to bacterial survival in the host cells.
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