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与肉孢子虫属18S rRNA可变区互补的寡核苷酸探针。

Oligonucleotide probes complementary to variable regions of 18S rRNA from Sarcocystis species.

作者信息

Holmdahl O J, Mattsson J G, Uggla A, Johansson K E

机构信息

Department of Cattle and Sheep Diseases, Swedish University of Agricultural Sciences, Uppsala.

出版信息

Mol Cell Probes. 1993 Dec;7(6):481-6. doi: 10.1006/mcpr.1993.1071.

Abstract

Sarcocystis is a large genus of cyst-forming coccidian parasites in the phylum Apicomplexa (Protista). Stable RNA was extracted from cystozoites of Sarcocystis cruzi, S. tenella, S. fusiformis, S. gigantea and Toxoplasma gondii. The partial sequences of the small sub-unit ribosomal RNA (18S rRNA) were determined by direct RNA sequencing with reverse transcriptase. The rRNA sequences were computer aligned with the published partial sequence of T. gondii, and three oligonucleotides complementary to different variable regions of the 18S rRNA were synthesized. The three probes were end-labelled with 32P and tested in filter hybridization experiments. One of the probes designed to be Sarcocystis genus-specific, did not cross-hybridize to stable RNA from T. gondii. Two of the probes were designed to be species-specific for S. cruzi and S. tenella, and these probes hybridized specifically with their respective targets.

摘要

肉孢子虫属是顶复门(原生生物)中一大类形成包囊的球虫寄生虫。从克鲁斯肉孢子虫、柔嫩肉孢子虫、梭形肉孢子虫、巨型肉孢子虫和刚地弓形虫的包囊型虫体中提取稳定RNA。通过逆转录酶直接RNA测序确定小亚基核糖体RNA(18S rRNA)的部分序列。将rRNA序列与已发表的刚地弓形虫部分序列进行计算机比对,并合成了与18S rRNA不同可变区互补的三种寡核苷酸。这三种探针用32P进行末端标记,并在滤膜杂交实验中进行测试。设计为肉孢子虫属特异性的一种探针未与刚地弓形虫的稳定RNA发生交叉杂交。另外两种探针设计为克鲁斯肉孢子虫和柔嫩肉孢子虫的种特异性探针,这些探针分别与各自的靶标发生特异性杂交。

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