Amplification and sequence of eastern Zhejiang white geese glyceraldehyde-3-phophate dehydrogenase for real time PCR applications

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a glycoytic enzyme, which is active in all vertebrate tissues and frequently used as a housekeeping gene in expression studies. In this study, the mRNA sequence of glyceraldehyde-3-phosphate dehydrogenase in duck was analyzed by designing a pair of primers and ampified in eastern Zhejiang white geese (a native Chinese geese breed) as a control gene in real time PCR. The mRNA levels of the target gene were measured relatively to their respective templates and were normalized to reference genes, to control the amount of cDNA loaded into the reaction. The sequences of genomic DNA and cDNA of goose glyceraldehydes-3-phophate dehydrogenase were 235bp and 209bp, respectively. Real time PCR showed a specific Ct using cDNA as template, and the primers showed a single melt peak. So the gene expression study was reliable when glyceraldehyde-3-phosphate dehydrogenase was used as an internal reference gene.