用于禽白血病病毒J亚群感染的鸡胚成纤维细胞定量实时PCR分析的内参基因选择
Selection of reference genes for quantitative real-time PCR analysis in chicken embryo fibroblasts infected with avian leukosis virus subgroup J.
作者信息
Yang Falong, Lei Xiaowen, Rodriguez-Palacios Alexander, Tang Cheng, Yue Hua
机构信息
Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China.
出版信息
BMC Res Notes. 2013 Oct 7;6:402. doi: 10.1186/1756-0500-6-402.
BACKGROUND
The selection of stably expressed reference genes is a prerequisite when evaluating gene expression, via real-time PCR, in cells in response to viral infections. The objective of our study was to identify suitable reference genes for mRNA expression analysis in chicken embryonic fibroblasts (CEF) after infection with avian leukosis virus subgroup J (ALV-J).
FINDINGS
The expression levels of 11 potential reference genes in CEF infected with ALV-J were determined by real-time PCR. The expression stability of these genes were analyzed and ranked using the geNorm tool. Analysis indicated that the genes RPL30 (ribosomal protein L30) and SDHA (succinate dehydrogenase complex, subunit A) were the most stably expressed genes in the ALV-J infected CEF.
CONCLUSIONS
The RPL30 and SDHA were deemed suitable for use as reference genes for real-time PCR analysis of mRNA gene expression during ALV-J infection, whereas commonly used ACTB and GAPDH are unsuitable to be reference genes.
背景
在通过实时定量PCR评估细胞对病毒感染的反应中的基因表达时，选择稳定表达的内参基因是一个先决条件。我们研究的目的是鉴定在感染禽白血病病毒J亚群（ALV-J）后鸡胚成纤维细胞（CEF）中用于mRNA表达分析的合适内参基因。
研究结果
通过实时定量PCR测定了感染ALV-J的CEF中11个潜在内参基因的表达水平。使用geNorm工具分析并排列了这些基因的表达稳定性。分析表明，基因RPL30（核糖体蛋白L30）和SDHA（琥珀酸脱氢酶复合物亚基A）是在感染ALV-J的CEF中表达最稳定的基因。
结论
RPL30和SDHA被认为适合用作ALV-J感染期间mRNA基因表达实时定量PCR分析的内参基因，而常用的ACTB和GAPDH不适合作为内参基因。
Zotero 插件