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苹果蠹蛾幼虫中转座子TCp3.2转座酶cDNA的序列分析与定量

Sequence analysis and quantification of transposase cDNAs of transposon TCp3.2 in Cydia pomonella larvae.

作者信息

Arends Hugo M, Jehle Johannes A

机构信息

Department of Phytopathology, Laboratory for Biotechnological Crop Protection, Agricultural Service Center Palatinate (DLR Rheinpfalz), Breitenweg 71, 67435 Neustadt an der Weinstrasse, Germany.

出版信息

Arch Insect Biochem Physiol. 2006 Nov;63(3):135-45. doi: 10.1002/arch.20149.

Abstract

The Tc1-like transposable element TCp3.2 was previously found to be horizontally transferred from the genome of Cydia pomonella to the C. pomonella granulovirus (CpGV). In this study, the transcription of transposase genes of endogenous TCp3.2 copies in the insect host genome was investigated. Cloning and sequencing of cDNAs prepared from TCp3.2 transposase transcripts resulted in the identification of a 199-bp-long intron. Sequence heterogeneities among different cDNA clones suggested that multiple copies of the transposase are transcribed, but that a part of these copies encode a defective transposase. The actin gene of C. pomonella was cloned and sequenced, and used to standardise quantitative real time PCR on prepared cDNA of the TCp3.2 transposase. Comparison of cDNA levels of TCp3.2 transposase prepared from mock and CpGV-infected C. pomonella larvae did not provide evidence that CpGV infection influenced the transcription level of TCp3.2 transposase.

摘要

此前发现,类Tc1转座元件TCp3.2从苹果蠹蛾基因组水平转移至苹果蠹蛾颗粒体病毒(CpGV)基因组中。在本研究中,对昆虫宿主基因组内源性TCp3.2拷贝的转座酶基因转录情况进行了研究。对由TCp3.2转座酶转录本制备的cDNA进行克隆和测序,鉴定出一个长度为199 bp的内含子。不同cDNA克隆之间的序列异质性表明,转座酶的多个拷贝被转录,但其中一部分拷贝编码有缺陷的转座酶。克隆并测序了苹果蠹蛾的肌动蛋白基因,并将其用于对制备的TCp3.2转座酶cDNA进行定量实时PCR标准化。比较从模拟感染和CpGV感染的苹果蠹蛾幼虫制备的TCp3.2转座酶cDNA水平,未发现CpGV感染影响TCp3.2转座酶转录水平的证据。

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