山羊不同组织器官的内参基因筛选

Screening of Internal Reference Genes in Different Tissues and Organs of Goats

本研究通过比较9个内参基因在山羊不同组织中的表达水平进而确定最适合研究山羊组织表达的内参基因。本试验以简州大耳羊为试验材料,利用实时荧光定量PCR技术分析9个内参基因(GAPDH,PPIA,18S rRNA,PPIB,UXT,RPLP0,ACTB,EIF3K和TBP)在心脏、肝脏、脾脏、肺脏、肾脏、大肠、瘤胃、背最长肌和皮下脂肪等组织中的表达差异情况,并利用geNorm、NormFinder和BestKeeper等程序分析了它们的表达稳定性。geNorm和NormFinder程序一致显示TBP表达最稳定,其次是UXT和RPLP0;BestKeeper分析显示18S rRNA表达最为稳定,其次为TBP和ACTB;3个程序一致认为GAPDH表达稳定性最差。综合3个程序分析得出TBP最适合作为山羊组织中的内参基因,其次为UXT和RPLP0,GAPDH表达稳定性最差,不适合作为山羊组织内参,这为后续研究其他目的基因在山羊组织器官中的表达模式提供数据保障。

In this study,the expression levels of nine internal reference genes in different tissues of goats were compared to determine the most suitable internal reference genes for the study of goat tissue expression.The study analyzed the expression differences of nine traditional reference genes(GAPDH,PPIA,18 S rRNA,PPIB,UXT,RPLP0,ACTB,EIF3 K and TBP)in Jianzhou Big-eared goat by real-time fluorescent quantitative PCR.Their expression stability was analyzed by ge Norm,Normfinder,and BestKeeper.Among them,geNorm and NormFinder consistently showed that TBP expression was the stablest,followed by UXT and RPLP0;BestKeeper analysis showed that 18 S rRNA expression was the stablest followed by TBP and ACTB;the three programs agreed that the stability of GAPDH expression was the worst.Comprehensive analysis of the three procedures showed that TBP expression was the most stable and most suitable for the reference gene in Jianzhou Big-eared goat tissue,followed by UXT and RPLP0,and GAPDH expression stability was not suitable as the internal reference of Jianzhou Big-eared sheep.This study provides data protection for the follow-up study of the expression patterns of other target genes in the Jianzhou Big-eared goat tissues.