畜牧兽医学报2018，Vol.49Issue(5)：907-918,12.DOI:10.11843/j.issn.0366-6964.2018.05.005
山羊肌内前体脂肪细胞诱导分化过程中内参基因的表达稳定性分析
The Expression Stability Analysis of Reference Genes in the Process of Goat Intramuscular Preadipocytes Differentiation in Goat
摘要
Abstract
The aim of this study was to determine the most suitable reference genes in different periods of goat intramuscular preadipocytes differentiation in goat by comparing the expression levels of 9 candidate reference genes.The intramuscular preadipocytes of goat were obtained by collagenase digestion technique.The adipocytes induced for 0,1,2,3,5 and 6 d were used in this study.The expression levels of the 9 candidate reference genes (GAPDH,PPIA,18S rRNA,PPIB,UXT,RPLP0,ACTB,EIF3K and TBP) were detected by real-time quantitative PCR (qPCR),and the expression stability was evaluated by geNorm,NormFinder and BestKeeper programs,individually.Meanwhile,the suitability of the selected reference genes for normalization was further assessed by the expression of KLF3 and KLF12.The results showed that UXT was identified as the most suitable reference gene by both geNorm and NormFinder analysis,while the PPIB was identified as the most suitable reference gene by BestKeeper analysis.The combined use of UXT and PPIB as the reference genes resulted in the optimal selection for nor malization of analyzed genes in different periods of intramuscular preadipocytes differentiation in goat based on the analysis of geNorm,NormFinder and BestKeeper;Furthermore,the expression levels of KLF3 and KLF12 normalized by the most stable reference genes (UXT and PPIB,UXT) were significantly different from that normalized by the least stable ones (EIF3K,18S rRNA).In conclusion,the expression of UXT was the most stable and it was the most suitable reference genes during the differentiation of goat intramuscular preadipocyte in goat,meanwhile the combined use of UXT and PPIB was found to lead to a more accurate result for target genes normalization.These results provided a foundation for functional research of genes related to preadipocytes differentiation in goats.关键词
山羊/肌内前体脂肪细胞/内参基因/实时荧光定量PCRKey words
goat/intramuscular preadipocytes/reference gene/real-time quantitative PCR分类
农业科技引用本文复制引用
许晴,林森,朱江江,王永,林亚秋..山羊肌内前体脂肪细胞诱导分化过程中内参基因的表达稳定性分析[J].畜牧兽医学报,2018,49(5):907-918,12.基金项目
国家自然科学基金项目(31672395) （31672395）
四川省“十三五”畜禽育种攻关项目(2016NYZ0045) （2016NYZ0045）
中央高校基本科研业务费专项基金项目(2017NGJPY06) （2017NGJPY06）
