激光生物学报, 2017, 26 (4): 379, 网络出版: 2017-11-07
翘嘴鳜per1 mRNA表达量分析中采用的内参基因稳定性比较
Comparative Research on the Stability of Reference Genes Based on mRNA Expression Analysis of per1 in Siniperca chuatsi
翘嘴鳜 per1基因 GeNorm软件 内参基因 Siniperca chuatsi per1 qRT-PCR quantitative real-time PCR geNorm software reference genes
摘要
为筛选生物钟核心基因per1表达定量中的相对稳定性最好的内参基因, 本研究取翘嘴鳜成鱼心脏、肝脏、肾脏、脑、红肌、白肌、肠、眼和脾等九个组织为研究对象, 选取GAPDH、18S rRNA、β-actin、rps29、RPL13a、B2M和EF1a为内参基因, 采用实时荧光定量PCR(qRT-PCR)对per1基因mRNA表达水平进行检测分析。研究结果表明18S rRNA和GAPDH的平均稳定值M最低, 相对表达量最稳定。以18S rRNA和GAPDH为内参基因时分析发现per1基因表达量在肝脏中最高。本研究为在鱼类per1 mRNA表达检测过程中选用稳定的内参基因提供了实验和理论参考。
Abstract
For the screening the best reference gene of the biological clock core gene per1 expression in quantitative analysis, we compared the stability of the 7 reference genes (GAPDH, 18S rRNA, β-actin, rps29, RPL13a, B2M and EF1a) in heart, brain, kidney, liver, slow muscle, fast muscle, intestine, spleen and eye tissues of the adult Siniperca chuatsi. By using GeNorm software, the results showed that the GAPDH and 18S rRNA are the the most stable reference genes in 9 tissues. With 18S rRNA and GAPDH as reference gene analysis of the expression of per1 in the liver was the highest. This study for the detection of per1 mRNA levels in fish tissues and in the selection of one or more reference genes provide experimental and theoretical reference.
吴萍, 张方亮, 杨程涌, 张建社, 褚武英. 翘嘴鳜per1 mRNA表达量分析中采用的内参基因稳定性比较[J]. 激光生物学报, 2017, 26(4): 379. WU Ping, ZHANG Fangliang, YANG Chengyong, ZHANG Jianshe, CHU Wuying. Comparative Research on the Stability of Reference Genes Based on mRNA Expression Analysis of per1 in Siniperca chuatsi[J]. Acta Laser Biology Sinica, 2017, 26(4): 379.