飞蝗前肠最适内参基因筛选

Screening of optimal internal reference genes in the foregut of Locusta migratoria

为了筛选在不同发育天数的飞蝗5龄若虫前肠中及其前肠不同部位能够稳定表达的最适内参基因,选用β-肌动蛋白(β-actin)、延伸因子1-α(EF-1α)、3-磷酸甘油醛脱氢酶(GAPDH)、核糖体蛋白49(RP 49)和α-微管蛋白(α-tubulin)5个基因作为候选内参基因,利用实时荧光定量PCR技术(qRT-PCR)分析各候选内参基因的相对表达量,采用geNorm、NormFinder和Bestkeeper软件相结合,筛选出5龄飞蝗前肠不同部位和不同发育天数若虫前肠中的最适内参基因。结果表明,β-actin和α-tubulin为前肠不同部位最适内参基因组合,β-actin、α-tubulin和GAPDH为不同发育天数若虫前肠中最适内参基因组合。本文为飞蝗肠道关键靶基因分子特性及进一步生物学功能研究提供重要理论基础。

In order to screen the optimal internal reference genes that can be stably expressed in different segments of foregut and foregut of 5th-instar Locusta migratoria on different developmental days,five candidate genes,includingβ-actin,EF-1α,GAPDH,RP 49 andα-tubulin,were selected as internal reference genes.Real time quantitative PCR(qRT-PCR)was performed to analyze the relative expression of each candidate gene.The optimal internal reference genes were screened out by using geNorm,NormFinder and Bestkeeper software.The results showed thatβ-actin andα-tubulin was the most suitable gene combination for different parts of foregut,andβ-actin,α-tubulin and GAPDH were the most suitable internal reference genes for foregut of 5th-instar nymphs of L.migratoria on different developmental days.This study provides an important theoretical basis for further study of the molecular characteristics and biological functions of the key target genes in L.migratoria.