GENETIC AND BIOLOGICAL DETERMINANTS OF AVIAN TUMOR VIRUS SUSCEPTIBILITY
Location: Avian Disease and Oncology Laboratory
Title: In search of a suitable reference gene for normalization of gene expression in MDV-infected chicken cells
Submitted to: Avian Immunology Research Group Abstract
Publication Type: Abstract Only
Publication Acceptance Date: June 24, 2008
Publication Date: June 24, 2008
Citation: Heidari, M. 2008. In search of a suitable reference gene for normalization of gene expression in MDV-infected chicken cells [abstract]. Avian Immunology Research Group Conference Proceedings. 10th Avian Immunology Research Group Conference, June 24-27, 2008, Sea World Nara Resort, Queensland, Australia. Paper No. 18.
Marek’s disease (MD) is a contagious lymphoproliferative disease of domestic chickens caused by a highly cell-associated alpha-herpesvirus, MD virus (MDV). The choice of an appropriate housekeeping gene as an endogenous reference gene is an essential requirement for relative quantification of gene expression analysis. The suitability of a reference gene for normalization of target gene expression level in MDV-infected chicken cells has not been investigated previously. In this study we examined the reliability of 19 housekeeping genes in the spleen cells of MDV-infected chickens. Transcriptional activities of genes encoding ADAM, ALAS1, Albumin, GUSB, EF1alpha1, HMBS, HPRT1, PPIB, PPID, RPL19, RPLP0, SDHA, TBP, YWHAZ, UBC, beta2M, TUBB, GAPDH, and beta-actin were examined in age-matched infected and control birds. The 19 genes were ranked according to the stability of the expression levels based on the standard deviation of the cycle threshold and the software program geNorm. Out of the 19 genes examined RPL19 and RPLP0 were selected as the most reliable reference genes for normalization of gene expression analysis in MDV-infected spleen cells. Beta-actin and GAPDH, the two commonly used reference genes, ranked as the number 15 and 17 in stability of expression, respectively. In summary, both RPL19 and RPLP0 are recommended as the reference genes for relative quantification of gene expression profiling in MDV-infected chickens either as single gene or preferably in combination.