标题：Screening and validation of reference genes using in RT-qPCR for gene expression studies in Paederus fuscipes, a medically and agriculturally important insect
摘要：AbstractPaederus fuscipesis a medically and agriculturally important insect all over the world.P. fuscipescause not only a skin condition but is also an aggressive predator in different agro-ecosystems. Prior to performing RT-qPCR under a variety of conditions to investigate the function of target genes inP. fuscipes, it is essential to screen reference genes. However, noP. fuscipesreference gene(s) has yet been discovered. Using theRefFindersoftware package, which includesΔCt, geNorm, NormFinder,andBestKeeper,we evaluated the stability of seven housekeeping genes ofP. fuscipesunder five conditions (developmental stage, diet, temperature, tissue and sex). During the RT-qPCR analysis,geNormsoftware was used to evaluate pairwise variation in order to identify the most appropriate number of reference genes. The results revealed that for developmental stagesRPL-13, EF1Aandβ-tubulin; for sex-related experimentsβ-tubulinandActin, for tissue-related experimentsPRS-18, 18SandRPL-13; for temperature-related experimentsβ-tubulinandPRL-13and diet-related experiments,β-tubulinandPRS-18are suitable reference genes. Furthermore, the associatedHSP-70(as the reporter gene) expression patterns differed significantly when the three most stable and three least stable reference genes were selected in different temperature treatments. This is the first time that a complete list of standardized reference genes has been given forP. fuscipesto use in an RT-qPCR study, which could be helpful for potential functional studies of target genes inP. fuscipes.
