Abstract:

The identification of housekeeping genes in organisms is important for quantitative analysis of gene expression. To identify stable housekeeping genes in various tissues of the giant freshwater prawn Macrobrachium rosenbergii, the relative expression data in multiple tissues, including gill, eye stalk, heart, hepatopancreas, stomach, big claw muscle, abdominal muscle, testis, androgenic gland and ovary, was used to identify new candidate housekeeping genes for quantitative analysis of gene expression in various tissues. The results showed that 1 902 genes were obtained with average FPKM values above 5 in the 10 tissues. Based on the average variation coefficients of these genes, 10 candidate genes were screened out randomly with smaller average variation coefficients than the common housekeeping gene β-actin. Further selection with gene expression stability parameter (M value) showed that, the M value of the selected 10 candidate genes in 10 tissues were all smaller than that of β-actin. Among the 10 candidate genes, the RNF10 gene exhibited the smallest M value, while the S/TPP2A gene showed the biggest M value except the β-actin gene. Quantitative PCR test on RNF10, S/TPP2A and β-actin gene varified that the M values of RNF10, S/TPP2A were smaller than that of β-actin, and the RNF10 gene exhibited the best expression stability. Thus, it was feasible to screen out housekeeping genes by using transcriptome data, variability, and the RNF10 gene could be used as a housekeeping gene with good expression stability in 10 tissues of of Macrobrachium rosenbergii.

Key words: Macrobrachium rosenbergii, housekeeping gene, expression stability parameter