Abstract: Selection of a suitable reference gene is an important prerequisite for precise analysis of target gene expression by Real-time quantitative PCR(qPCR). In this paper, the partial sequence of three reference genes, β-actin and GAPDH, EF1-α, from spanish mackerel Scomberomorus niphonius were firstly cloned, and then the Ct values of the four reference genes in various tissues were measured by qPCR. Three different statistical algorithms including geNorm, NormFinder and BestKeeper were used to analyze the expression stability of the four reference genes in different tissues. The result showed that the most stable reference gene was EF1-a, followed by 18S ribosomal RNA. It provides a useful basis for selecting of the appropriate reference gene in Spanish Mackerel.

Key words: Scomberomorus niphonius, qPCR, reference gene, geNorm, expression stability