广东海洋大学学报2018，Vol.38Issue(1)：7-13,7.DOI:10.3969/j.issn.1673-9159.2018.01.002
光裸星虫全组织荧光定量PCR分析中内参基因的筛选
Screening of Reference Genes for Real-time PCR in Whole Tissue from Sipunculus nudus
摘要
Abstract
[Objective] To obtain the most stable reference gene in tissues of Sipunculus nudus (trunk muscle,intestinal,nephridium,introvert,retractor muscle,and coelomocytes).[Method] Quantitative Real-time PCR (qRT-PCR) was adopted to analyze eight traditional reference genes which including 3-phosphate dehydrogenase (GAPDH),β-actin,tubulin (TUB),TATA box binding protein (TBP),and ubiquitin C (UBC).Combining geNorm,NormFinder,BestKeeper and RefFinder software,the expression stability of eight reference genes had been analyzed.[Result] The results showed that specific amplification products could be obtained by the eight reference genes.The results of the stability ranking given by the four software were integrated by the cumulative integration method.The reference genes stability was descended as follows:TBP (1) ＞ TBP (2) ＞ UBC ＞ GAPDH＞ TUB (2) ＞TUB (1) ＞ β-actin (2) ＞β-actin (1).[Conclusion] The TBP (1) could be used as a suitable reference gene in qRT-PCR analysis of S.nudus.The results identified a suitable reference gene and laid the foundation for the follow-up research in the molecular biology.关键词
荧光定量PCR/内参基因/光裸星虫Key words
quantitative Real-time PCR/reference gene/Sipunculus nudus分类
生物科学引用本文复制引用
张家炜,郑哲,王庆恒,黄荣莲,杜晓东..光裸星虫全组织荧光定量PCR分析中内参基因的筛选[J].广东海洋大学学报,2018,38(1):7-13,7.基金项目
广东省科技计划(2016A020209010) （2016A020209010）
广东省自然科学基金(2017A030303075) （2017A030303075）
