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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제27권 제1호
- 발행연도
- 2017.1
- 수록면
- 112 - 121 (10page)
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초록· 키워드
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Edwardsiella tarda is a gram-negative pathogenic bacterium in aquaculture that can cause hemorrhagic septicemia in fish. Many secreted proteins have already been identified as virulent factors of E. tarda. Moreover, since virulent phenotypes are based on the expression regulation of virulent genes, understanding the expression profile of virulent genes is important. A quantitative RT-PCR is one of the preferred methods for determining different gene expressions. However, this requires the selection of a stable reference gene in E. tarda, which has not yet been systematically studied. Accordingly, this study evaluated nine candidate reference genes (recA, uup, rpoB, rho, topA, gyrA, groEL, rpoD, and 16S rRNA) using the Excel-based programs BestKeeper, GeNorm, and NormFinder under different culture conditions. The results showed that 16S rRNA was more stable than the other genes at different culture growth phases. However, at the same culture time, topA was identified as the reference gene under the conditions of different strains, different culture media, and infection, whereas gyrA was identified under the condition of different temperatures. Thus, in experiments, the expression of gapA and fbaA in E. tarda was analyzed by RT-qPCR using 16S rRNA, recA, and uup as the reference genes. The results showed that 16S rRNA was the most suitable reference gene in this analysis, and that using unsuitable reference genes resulted in inaccurate results.
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참고문헌 (20)
참고문헌 신청
Andersen CL / 2004 / Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normaliz
Blau K / 2007 / Flamingo cadherin: a putative host receptor for Streptococcus pneumoniae / J. Infect. Dis 195 : 1828 ~ 1837
Egea L / 2007 / Role of secreted glyceraldehyde-3-phosphate dehydrogenase in the infection mechanism of enterohemorrhagic and enteropathogenic Escherichia coli:interaction of
Henderson B / 2011 / Bacterial virulence in the moonlight: multitasking bacterial moonlighting proteins are virulence determinants in infectious disease / Infect. Immun 79 :
Huggett J / 2005 / Real-time RTPCR normalisation; strategies and considerations / Genes Immun 6 : 279 ~ 284
Blau K / 2007 / Flamingo cadherin: a putative host receptor for Streptococcus pneumoniae / J. Infect. Dis 195 : 1828 ~ 1837
Egea L / 2007 / Role of secreted glyceraldehyde-3-phosphate dehydrogenase in the infection mechanism of enterohemorrhagic and enteropathogenic Escherichia coli:interaction of
Henderson B / 2011 / Bacterial virulence in the moonlight: multitasking bacterial moonlighting proteins are virulence determinants in infectious disease / Infect. Immun 79 :
Huggett J / 2005 / Real-time RTPCR normalisation; strategies and considerations / Genes Immun 6 : 279 ~ 284
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