Validation of endogenous reference genes for insecticide-induced and developmental expression profiling of Liposcelis bostsrychophila (Psocoptera: Liposcelididae)
Validation of endogenous reference genes for insecticide-induced and developmental expression profiling of Liposcelis bostsrychophila (Psocoptera: Liposcelididae)
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验证 Liposcelis bostsrychophila（裸翅目：Liposcelididae）杀虫剂诱导和发育表达谱的内源参考基因
DOI：
10.1007/s11033-009-9803-0
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发表时间：
2010-02-01
影响因子：
2.8
通讯作者：
Wang, Jin-Jun
中科院分区：
文献类型：
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作者：
Jiang, Hong-Bo;Liu, Yong-Hua;Wang, Jin-Jun
采用逆转录- pcr （RT-PCR）和cDNA末端快速扩增（RACE）相结合的方法，从bostrychophila Liposcelis bostrychphila中克隆出4个管家基因Lb β - actin1、Lb β - actin2、Lb α - tubulin和LbGapdh。GenBank登录号分别为FJ196622、FJ447483、FJ595242、FJ595241。Lb β - actin1全长1772 bp，其中一个开放阅读框（ORF）编码376个氨基酸；Lb β - actin2全长1350 bp，包含一个开放阅读框（ORF）编码376个氨基酸。此外，Lb α - tubulin的1565 bp cDNA的ORF为1350 bp，编码450个氨基酸。LbGapdh具有333个氨基酸的ORF。序列分析和从编码区核苷酸序列生成的系统发育树显示，它们与其他昆虫的关系比与哺乳动物的关系更接近。对这4个基因与18S rRNA在乳杆菌中的转录稳定性进行定量分析，geNorm软件对不同发育阶段的乳杆菌转录稳定性排序从高到低依次为溴氰菊酯诱导的Lb β - actin1 > LbGapdh > Lb α -微管蛋白> Lb β - actin2 > Lb18S rRNA，对不同发育阶段的乳杆菌转录稳定性排序为Lb β - actin1 > Lb α -微管蛋白> Lb β - actin2 > LbGapdh > Lb18S rRNA。
Four housekeeping genes named Lb beta-Actin1, Lb beta-Actin2, Lb alpha-Tubulin, and LbGapdh were cloned from Liposcelis bostrychophila using the combined techniques of reverse transcriptase-PCR (RT-PCR) with rapid amplification of cDNA ends (RACE). The GenBank accession numbers were FJ196622, FJ447483, FJ595242, and FJ595241, respectively. The full-length cDNA of Lb beta-Actin1 was a 1,772 bp sequence with an open reading frame (ORF) encoded 376 amino acids, while Lb beta-Actin2 was 1,350 bp in length containing an ORF encoded 376 amino acids. Furthermore, the 1,565 bp cDNA of Lb alpha-Tubulin had an ORF of 1,350 bp encoding 450 amino acids. And LbGapdh possessed an ORF of 333 amino acids. Sequences analysis and phylogenetic trees generated from the nucleotide sequences of their coding regions revealed a relationship that was closer to other insects than to mammals. The four genes together with 18S rRNA were quantified for transcription stability in L. bostrychophila, and the geNorm software ranked from the most to least were Lb beta-Actin1 > LbGapdh > Lb alpha-Tubulin > Lb beta-Actin2 > Lb18S rRNA for deltamethrin induction, while ranked Lb beta-Actin1 > Lb alpha-Tubulin > Lb beta-Actin2 > LbGapdh > Lb18S rRNA for the different developmental stages.