Identification of normalization factors for quantitative real-time RT-PCR analysis of gene expression in Pacific abalone Haliotis discus hannai

摘要: Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is widely used in studies of gene expression. In most these studies, housekeeping genes are as internal references without validation. To identify appropriate reference for qRT-PCR Pacific abalone Haliotis discus hannai, we examined the transcription stability six tissues presence and absence bacterial infection. For this purpose, were infected with pathogen Vibrio anguillarum 12 h 48 h. The mRNA levels five (digestive glands, foot muscle, gill, hemocyte, mantle) determined by qRT-PCR. PCR data was subsequently analyzed geNorm NormFinder algorithms. results show that infection, elongation factor-1-alpha beta-actin stably expressed all tissues, thus suitable cross-tissue type normalization factors. However, did not any universal post infection because stable varied between tissue types. Furthermore, optimal identified both algorithms at post-infection differed. These indicate induced significant changes expression a manner dependent on duration As result, different factors must be points.