摘要
目的反转录聚合酶链反应（RT—PCR）广泛应用于基因表达分析，其中选择合适的管家基因非常重要。大鼠心肌梗死模型是心肌梗死相关研究的主要动物模型，而心肌梗死后大鼠管家基因选择问题研究报道则较少。本研究拟探讨大鼠心梗模型中管家基因表达的稳定性。方法结扎冠脉前降支制作心梗模型，分析并挑选出使用广泛的4个标准管家基因：3-磷酸甘油醛脱氢酶（GAPDH），核糖体蛋白L13A（RPL13A），β-肌动蛋白（ACTB），结合区连接蛋白（ARBP）；用实时荧光反转录-聚合酶链反应方法比较它们在心梗大鼠心脏中的表达，采用GeNorm程序分析最适于心肌梗死后大鼠基因表达管家基因研究。结果RPL13A、GAPDH、ARBP及ACTB基因表达稳定度的平均值（M值）分别为0．812、0．721、0．812和1．2。结论GAPDH及ARBP是大鼠心梗模型中表达最稳定的基因。
Objective Reverse transcription-polymerase chain reaction （RT-PCR） is widely used in gene expression analysis.Selection of proper housekeeping gene is very important. Rat model of myocardial infarction is a major animal model of myocardial infarction. Few reports are found about the selection of rat housekeeping gene after myocardial infarction. This study was aimed to explore the stability of the housekeeping genes expression in the rat model of myocardial infarction. Methods Myocardial infarction models were made using the anterior descending coronary artery ligation method. Combining with practical work, analysis and selection were made of four widely used standard housekeeping genes： glyceraldehydes-3-phosphate dehydrogenase （GAPDH）, ribosomal protein L13A （RPL13A）, beta-actin （ACTB） and acidic ribosomal phosphoprotein P0 （ARBP）. The expressions in the rat heart were compared using real-time fluorescent RT-PCR and analysis was carried out to figure out which was the most suitable housekeeping gene for study of gene expression in rats after myocardial infarction using GeNorm program. Results The M values of RPL13A, GAPDH, ARBP and ACTB gene were 0.812, 0.721, 0.812 and 1.2 respectively. Conclusion GAPDH and ARBP are the most stable genes for the rat myocardial infarction model.
出处
《国际生物医学工程杂志》
CAS
2013年第4期216-219,I0004,I0005,共6页
International Journal of Biomedical Engineering
