桔小实蝇不同组织基因表达转录分析中内参基因的筛选

In this study, we investigated the stability of ten candidate reference genes for gene expression study in three different tissues (midgut, Malpighian tubules, and fat body) of the oriental fruit fly, Bactrocera dorsalis (Hendel). Two different programs, geNorm and Normfinder, were used to analyze the data. According to geNorm, α-TUB + ACT5 are the most appropriate reference genes for gene expression profiling across the three different tissues in the female flies, while ACT1 + ACT2 are considered the best for males. Furthermore, we evaluated the stability of the reference gene candidates, and selected out appropriate reference gene to determine the sexual differences in gene expression in the same tissue. In the midgut and Malpighian tubules, ACT2 + ACT3 are the best choice for both males and females. However, α-TUB + ACT1 are the best pair for fat body. Although the results calculated by Normfinder are different from geNorm in rankings, α-TUB is always one of the most stable genes in each sample calculated by the two programs. This work not only formed a solid basis for future gene expression study using real-time PCR, but also will serve as a resource to screen insect reference genes for gene expression studies in any other insects.