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捻转血矛线虫:一种两步差异显示PCR技术的开发,用于检测免疫羊和未免疫羊体内线虫的差异基因表达

Haemonchus contortus: development of a two-step, differential-display PCR to detect differential gene expression in nematodes from immune and naïve sheep.

作者信息

Rowe A, Yun K, Emery D, Sangster N

机构信息

Faculty of Veterinary Science, McMaster Building (B14), University of Sydney, Camperdown Campus, NSW 2006, Australia.

出版信息

Exp Parasitol. 2008 Jun;119(2):207-16. doi: 10.1016/j.exppara.2008.01.015. Epub 2008 Feb 7.

Abstract

Haemonchus contortus is a blood-feeding nematode which parasitizes the abomasum of sheep and represents a serious constraint to sheep production. Anthelmintics are currently the most common method of worm control but the worldwide development of multiple-drug resistance and issues of residues in the food chain make alternatives to anthelmintics a priority. Biotechnology-driven solutions to parasitism include vaccines and silencing of genes regulating nematode development. To pursue gene targets that may be suitable for parasite control, a two stage differential-display PCR (dd-PCR) approach was developed to observe differential gene expression between Haemonchus from immune and control sheep. Twenty-four reproducible differentially-expressed bands were identified in 60 pairs of dd-PCR comparisons. The first of these cloned and sequenced corresponded to the H. contortus 60S ribosomal protein L35A. The remaining bands are being cloned and validated and may provide new targets for parasite control.

摘要

捻转血矛线虫是一种吸食血液的线虫,寄生于绵羊的皱胃,对绵羊生产构成严重制约。驱虫药是目前控制蠕虫最常用的方法,但全球范围内多重耐药性的发展以及食物链中的残留问题使得驱虫药的替代方法成为当务之急。生物技术驱动的寄生虫防治解决方案包括疫苗和调节线虫发育的基因沉默。为了寻找可能适用于寄生虫控制的基因靶点,开发了一种两阶段差异显示PCR(dd-PCR)方法,以观察免疫绵羊和对照绵羊体内捻转血矛线虫之间的差异基因表达。在60对dd-PCR比较中鉴定出24条可重复的差异表达条带。其中第一条克隆并测序的条带对应于捻转血矛线虫60S核糖体蛋白L35A。其余条带正在克隆和验证,可能为寄生虫控制提供新的靶点。

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