Reference Genes For Expression Analysis Research Articles

Housekeeping genes for quantitative expression studies in the three-spined stickleback Gasterosteus aculeatus

BackgroundDuring the last years the quantification of immune response under immunological challenges, e.g. parasitation, has been a major focus of research. In this context, the expression of immune response genes in teleost fish has been surveyed for scientific and commercial purposes. Despite the fact that it was shown in teleostei and other taxa that the gene for beta-actin is not the most stably expressed housekeeping gene (HKG), depending on the tissue and experimental treatment, the gene has been used as a reference gene in such studies. In the three-spined stickleback, Gasterosteus aculeatus, other HKG than the one for beta-actin have not been established so far.ResultsTo establish a reliable method for the measurement of immune gene expression in Gasterosteus aculeatus, sequences from the now available genome database and an EST library of the same species were used to select oligonucleotide primers for HKG, in order to perform quantitative reverse-transcription (RT) PCR. The expression stability of ten candidate reference genes was evaluated in three different tissues, and in five parasite treatment groups, using the three algorithms BestKeeper, geNorm and NormFinder. Our results showed that in most of the tissues and treatments HKG that could not be used so far due to unknown sequences, proved to be more stably expressed than the one for beta-actin.ConclusionAs they were the most stably expressed genes in all tissues examined, we suggest using the genes for the L13a ribosomal binding protein and ubiquitin as alternative or additional reference genes in expression analysis in Gasterosteus aculeatus.

Zebrafish GAPDH can be used as a reference gene for expression analysis in cross-subfamily cloned embryos

Nuclear transfer between two Wsh species oVers a usefultechnique to study nucleo-cytoplasmic interaction andnuclear reprogramming [1,2]. Recently, we generated cross-genus cloned Wsh derived from common carp nuclei andgoldWsh enucleated eggs and found that the somitogenesisand vertebral numbers of cloned Wsh were consistent to theegg-providing species goldWsh, but not to the donor cellspecies common carp [2]. Therefore, it was of great impor-tance to investigate the diVerence of gene expression pat-tern in interspecies cloned Wsh compared with cell donorspecies. Cross-subfamily nuclear transfer between two labo-ratory Wsh species, rare minnow (