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奥斯特他线虫属、血矛线虫属、古柏线虫属和食道口线虫属:用于区分牛重要寄生虫的属特异性DNA探针的构建与特性分析

Ostertagia, Haemonchus, Cooperia, and Oesophagostomum: construction and characterization of genus-specific DNA probes to differentiate important parasites of cattle.

作者信息

Christensen C M, Zarlenga D S, Gasbarre L C

机构信息

Helminthic Diseases Laboratory, USDA-ARS, Beltsville, Maryland 20705.

出版信息

Exp Parasitol. 1994 Feb;78(1):93-100. doi: 10.1006/expr.1994.1009.

Abstract

Partial genomic DNA libraries for four common cattle parasites, Ostertagia ostertagi (Oo), Haemonchus placei (Hp), Cooperia oncophora (Co), and Oesophagostomum radiatum (Or), were generated and differentially screened with radiolabeled homologous and heterologous genomic DNA. Clones were identified from each parasite species which did not cross react with DNA from the heterologous genera by Southern blot or slot-blot analyses. Four clones, pOo2, pHp2.1, pCo2, and pOr14 were chosen for further examination. Genomic DNA from other species within the genera were screened with the cloned segments, and the clones were shown to be genus specific. The cloned fragments ranged in size from 195 to 669 base pairs and were approximately 50% in AT content. The cloned segments, pOo2 and pCo2, were approximately equally represented within their parasite genomes at 0.40 and 0.62%, respectively, whereas pHp2.1 and pOr14 were significantly higher at 1.61 and 3.39%, respectively. Sequence analysis demonstrated that pOr14 contained three 180-base pair tandemly repeated units with minor sequence variability between the repeats. Results presented here suggest that these sequences can detect as few as 25 eggs, and hence, may be used as diagnostic probes for the antemortem differentiation of trichostrongyle infections in cattle.

摘要

构建了四种常见牛寄生虫——奥斯特他线虫(Oo)、柏氏血矛线虫(Hp)、牛库珀线虫(Co)和辐射食道口线虫(Or)的部分基因组DNA文库,并用放射性标记的同源和异源基因组DNA进行差异筛选。通过Southern印迹或狭缝印迹分析,从每个寄生虫物种中鉴定出不与异源属DNA发生交叉反应的克隆。选择了四个克隆,即pOo2、pHp2.1、pCo2和pOr14进行进一步研究。用克隆片段筛选该属内其他物种的基因组DNA,结果表明这些克隆具有属特异性。克隆片段的大小在195至669个碱基对之间,AT含量约为50%。克隆片段pOo2和pCo2在其寄生虫基因组中的占比分别约为0.40%和0.62%,而pHp2.1和pOr14的占比则显著更高,分别为1.61%和3.39%。序列分析表明,pOr14包含三个180个碱基对的串联重复单元,重复序列之间的序列变异较小。此处给出的结果表明,这些序列能够检测低至25个虫卵,因此可作为牛毛圆线虫感染生前鉴别诊断的探针。

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