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天然存在的克氏锥虫变体的鉴定、分离及特性分析。

Identification, isolation, and characterization of naturally-occurring Trypanosoma cruzi variants.

作者信息

McDaniel J P, Dvorak J A

机构信息

Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.

出版信息

Mol Biochem Parasitol. 1993 Feb;57(2):213-22. doi: 10.1016/0166-6851(93)90197-6.

Abstract

Naturally occurring DNA variants of the single-cell-derived Y-02 stock of Trypanosoma cruzi were discovered during a routine assay of the stock. Three DNA variant types were isolated. One type was indistinguishable from the parental Y-02 stock on the basis of total DNA cell-1. The other two types contained approximately 30% and 70% more DNA cell-1 than the parental Y-02 stock. Both the nucleus and kinetoplast were involved in the DNA content differences. The increase in DNA cell-1 was not G-C- or A-T-specific and was unrelated to the developmental stage of the parasite. Epimastigote population doubling times, isoenzymes, and schizodeme analyses could not differentiate the variant stocks. However, marked karyotype polymorphisms were observed by pulse-field gel electrophoresis, and restriction-fragment-length-polymorphisms were detected in hybridizations of some endonuclease-restricted samples to the spliced leader probe. We postulate that the Y-02 variants are genetic homologs. The ability to form viable hybrids or aneuploids provides T. cruzi with a mechanism to survive environmental stress, promote intra-specific heterogeneity and generate the diversity observed in the presentation and course of Chagas' disease.

摘要

在对克氏锥虫单细胞衍生的Y-02虫株进行常规检测时,发现了其天然存在的DNA变异体。分离出了三种DNA变异体类型。其中一种类型基于每个细胞的总DNA与亲本Y-02虫株无法区分。另外两种类型每个细胞的DNA比亲本Y-02虫株分别多约30%和70%。细胞核和动基体都与DNA含量差异有关。每个细胞DNA的增加并非特定于G-C或A-T,且与寄生虫的发育阶段无关。前鞭毛体群体倍增时间、同工酶和裂殖体分析无法区分变异虫株。然而,通过脉冲场凝胶电泳观察到明显的核型多态性,并且在一些经核酸内切酶切割的样品与剪接前导探针的杂交中检测到限制性片段长度多态性。我们推测Y-02变异体是遗传同源物。形成有活力的杂种或非整倍体的能力为克氏锥虫提供了一种在环境压力下生存、促进种内异质性以及产生恰加斯病表现和病程中所观察到的多样性的机制。

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