Selection Of Reference Genes From Different Development Periods And Tissues In Zi Geese

The purpose of the present study was to detect the expression of candidate reference genes 28S rRNA, 18S rRNA, GAPDH, ACTB, HPRT1, SDH and TUB in 1 day old, 2 months old, 4 months old, 6 months old and 8 months old Zi goose's liver, kidney, heart, muscle and ovary by qRT-PCR. Then the expression stability of these genes was analysed by geNorm and NormFinder software, finally the most appropriate reference genes and their numbers were screened.This study detect 7 candidate reference genes of 1 day old,2 months old,4 months old,6 months old and 8 months old Zi goose'tissues. After analysed by geNorm,the stability of genes were ranked from high to low according to genes according the stability value (M). The specific sequencing were as follows. 1 day old: GAPDH/HPRT1>28S rRNA>TUB>SDH>ACTB>18S rRNA; 2 months old: HPRT1/28S rRNA>GAPDH>18S rRNA>ACTB>TUB>SDH; 4 months old: GAPDH /28S rRNA>HPRT1>SDH>18S rRNA>ACTB>TUB; 6 months old: GAPDH / HPRT1>TUB>28S rRNA >18S rRNA>ACTB>SDH; 8 months old: GAPDH/28S rRNA>ACTB>TUB>HPRT1>SDH>18S rRNA. And V2/3 of pairwise variance analysis of 7 candidate reference genes were separately 0.129, 0.051, 0.080, 0.084, 0.069. The appropriate number of reference genes was 2. Meanwhile, relative steady reference genes that were screened by NormFinder were separately GAPDH and HPRT1 (1 day old), HPRT1 and 28S rRNA (2 months old), GAPDH and 28S rRNA (4 months old), GAPDH and HPRT1 (6 months old), GAPDH and 28S rRNA (8 months old),these results were as same as results of geNorm.This study detect 7 candidate reference genes of liver, kidney, heart, muscle and ovary at different development stages in by qRT-PCR in Zi goose ,the stability of genes were ranked from high to low after analysed by geNorm.The specific sequencing were as follows. heart:GAPDH/ 28S rRNA > SDH >ACTB>18S rRNA>HPRT1>TUB, liver: GAPDH/HPRT1>SDH>TUB>28S rRNA>18S rRNA>ACTB, kidney: ACTB/SDH>GAPDH>HPRT1>28S rRNA>18S rRNA>TUB; muscle: HPRT1/18S rRNA> GAPDH>28S rRNA>SDH>ACTB>TUB; ovary: GAPDH/HPRT1>SDH>TUB>18S rRNA>ACTB >28S rRNA. And pairwise variance analysis of 7 candidate reference genes were separately heart: V4/5= 0.210, liver: V2/3=0.237, kidney: V3/4=0.148, muscle: V2/3=0.124, ovary: V2/3=0.152. The appropriate number of reference genes was 4, 2, 3, 2 and 2. Meanwhile, relative steady reference genes that were screened by NormFinder were separately 28S rRNA, SDH, GAPDH and HPRT1(heart), GAPDH and HPRT1(liver), ACTB, SDH and GAPDH(kidney), HPRT1 and 18S rRNA(muscle), GAPDH and HPRT1(ovary), these results were as same as results of geNorm .