实时荧光定量PCR研究蛋鸡组织基因表达的内参基因筛选

发布时间：2018-05-20 05:13

本文选题：蛋鸡 + 内参基因　；参考：《中国畜牧兽医》2017年08期

【摘要】：为筛选出蛋鸡不同组织中稳定表达的内参基因,试验以120日龄的海兰灰蛋鸡(Gallus domesticus)为试验动物,选取常见内参基因RPS2、β-actin、GAPDH和HMBS作为候选基因,利用实时荧光定量PCR技术Ct值分析法对4个候选内参基因的相对表达进行测定,利用独立评价软件geNorm和NormFinder对蛋鸡的4个候选内参基因在不同组织(心脏、肝脏、肺脏、肾脏、肠、胫骨、胰脏和子宫)中的表达稳定性进行评价。结果显示,Ct值分析和geNorm程序分析均得出相似的结果,即RPS2基因始终占主导地位,稳定性最强,β-actin基因次之;NormFinder软件分析显示,HMBS基因稳定性最好,最佳组合为HMBS和RPS2基因;当分析不同组织中表达恒定的内参基因时,发现不同组织最稳定的内参基因也不完全相同。由此说明,不同的组织器官和不同的分析方法得出的结论虽然不完全一致,但它们却有着一定的潜在共性,发展趋势有一定相似性,即RPS2和β-actin基因相对稳定性更强。
[Abstract]:In order to screen the stable expression of internal reference genes in different tissues of laying hens, 120 days old Hailan gray laying hens (Gallus domesticusus) were used as experimental animals. The common internal reference genes RPS2, 尾 -actinine GAPDH and HMBS were selected as candidate genes. The relative expression of four candidate internal reference genes was determined by real-time fluorescence quantitative PCR assay. The four candidate internal reference genes were detected in different tissues (heart, liver, lung, kidney) by independent evaluation software geNorm and NormFinder. Expression stability was evaluated in intestine, tibia, pancreas and uterus. The results showed that the results of Ct value analysis and geNorm program analysis showed that the RPS2 gene was always dominant and the stability was the strongest, and the 尾 -actin gene followed by Norm Finder software analysis showed that the stability of HMBS gene was the best, and the best combination was HMBS and RPS2 gene. It was found that the most stable internal reference genes were not the same in different tissues when they were expressed in different tissues. It shows that although the conclusions of different tissues and organs and different analytical methods are not completely consistent, they have certain potential commonness, and the development trend has certain similarity, that is, the relative stability of RPS2 and 尾 -actin genes is stronger than that of 尾 -actin gene.
【作者单位】：贵州大学动物科学学院;高原山地动物遗传育种与繁殖教育部重点实验室;
【基金】：国家自然科学基金(31560642)
【分类号】：S831

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