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盘尾丝虫两个重复DNA序列的克隆与特性分析

Cloning and characterization of two Onchocerca volvulus repeated DNA sequences.

作者信息

Perler F B, Karam M

出版信息

Mol Biochem Parasitol. 1986 Nov;21(2):171-8. doi: 10.1016/0166-6851(86)90020-4.

Abstract

Two repeated sequences, plasmids pOV8 and pOV26, were cloned and characterized from the filarial parasite Onchocerca volvulus. Both clones can be used to distinguish O. volvulus DNA from other Onchocerca species or other nematodes by restriction fragment length polymorphisms, but neither clone can differentiate between DNA from savanna (Mali) or forest (Ivory Coast) O. volvulus isolates. DNA from one O. volvulus infective larva can be detected by both clones in dot blot hybridization assays. Neither clone cross hybridizes with DNA from host or vector species (human or simuliid, respectively). pOV26 is a member of an interspersed repeated DNA family composed of at least 100 members, and is only observed in the genus Onchocerca. Repeated DNA clone pOV8 cross reacts with DNA from other parasitic filarial nematodes, and is also present in at least 100 copies per O. volvulus genome. pOV26 is a potential tool in the diagnosis of human onchocerciasis, since it is specific for the genus Onchocerca. In the future, we plan to look for regions of these repeated sequences which may serve as a basis for the development of probes to discriminate among Onchocerca species and strains using a simple dot hybridization test.

摘要

从丝虫寄生虫旋盘尾丝虫中克隆并鉴定了两个重复序列,即质粒pOV8和pOV26。通过限制性片段长度多态性,这两个克隆都可用于区分旋盘尾丝虫的DNA与其他盘尾丝虫物种或其他线虫的DNA,但两个克隆都无法区分来自稀树草原(马里)或森林(象牙海岸)的旋盘尾丝虫分离株的DNA。在斑点印迹杂交试验中,两个克隆都能检测到来自一只旋盘尾丝虫感染性幼虫的DNA。两个克隆均不与宿主或载体物种(分别为人或蚋)的DNA交叉杂交。pOV26是一个散布重复DNA家族的成员,该家族至少由100个成员组成,且仅在盘尾丝虫属中观察到。重复DNA克隆pOV8与其他寄生性丝虫线虫的DNA发生交叉反应,并且每个旋盘尾丝虫基因组中也至少存在100个拷贝。pOV26是诊断人类盘尾丝虫病的潜在工具,因为它对盘尾丝虫属具有特异性。未来,我们计划寻找这些重复序列的区域,这些区域可能作为开发探针的基础,以便使用简单的斑点杂交试验来区分盘尾丝虫的物种和菌株。

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