原创力文档an evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either piscirickettsia salmonis or ipnv评估潜在的参考基因稳定表达在感染后两salmonid细胞系piscirickettsia salmonis或ipnv.pdfVIP
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an evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either piscirickettsia salmonis or ipnv评估潜在的参考基因稳定表达在感染后两salmonid细胞系piscirickettsia salmonis或ipnv
Peña et al. BMC Research Notes 2010, 3:101
/1756-0500/3/101
SHORT REPORT Open Access
An evaluation of potential reference genes for
stability of expression in two salmonid cell lines
after infection with either Piscirickettsia salmonis
or IPNV
1* 2 1
Andrea A Peña , Niels C Bols , Sergio H Marshall
Abstract
Background: Due to the limited number of species specific antibodies against fish proteins, differential gene
expression analyses are vital for the study of host immune responses. Quantitative real-time reverse transcription
PCR (qRT-PCR) is one of the most powerful tools for this purpose. Nevertheless, the accuracy of the method will
depend on the careful selection of genes whose expression are stable and can be used as internal controls for a
particular experimental setting.
Findings: The expression stability of five commonly used housekeeping genes [beta-actin (ACTB), elongation factor
1-alpha (EF1A), ubiquitin (UBQ), glyceraldehyd-3-phosphate dehydrogenase (GAPDH) and tubulin alpha (TUBA)] were
monitored in salmonid cell lines CHSE-214 and RTS11 after infection with two of the most fastidious fish
pathogens, the facultative bacterium Piscirickettsia salmonis and the aquabirnavirus IPNV (Infectious Pancreatic
Necrosis Virus). After geNorm analysis, UBQ and EF1A appeared as the most stable, although EF1A was slightly
upregulated at late stages of P. salmonis infection in RTS11. ACTB instead, showed a good performance in each
case, being always considered within the three most stable genes of the panel. In contrast, infection-dependent
differential regulation of GAPDH and TUBA was also demonstrated.
Conclusion: Based on the data presented here with the cell culture models CHSE-214 and RTS11, we suggest the
initial choice of UBQ, ACTB and EF1
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