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在指数富集的配体系统进化(SELEX)过程中使用胶体金筛选针对KMP-11的适体。

Selection of aptamers against KMP-11 using colloidal gold during the SELEX process.

作者信息

Moreno Miguel, Rincón Esther, Piñeiro David, Fernández Gerónimo, Domingo Alberto, Jiménez-Ruíz Antonio, Salinas Matilde, González Víctor M

机构信息

Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Alcalá, 28871 Alcalá de Henares, Madrid, Spain.

出版信息

Biochem Biophys Res Commun. 2003 Aug 22;308(2):214-8. doi: 10.1016/s0006-291x(03)01352-4.

Abstract

SELEX procedure is a methodology in which single stranded oligonucleotides are selected from a wide variety of sequences based on their interaction with a target molecule. We have designed a novel SELEX methodology using colloidal gold to select high affinity single stranded DNA aptamers against Leishmania infantum KMP-11. Kinetoplastid membrane protein-11 (KMP-11) is a major component of the cell membrane of kinetoplastid parasites. Although its function is not known, the fact that KMP-11 is a cytoskeleton-associated protein suggests that it may be involved in mobility or in some other aspects of the flagellar structure. We have isolated a single stranded DNA aptamer population that binds specifically to L. infantum KMP-11. This population has been characterized in a series of in vitro experiments suggesting that it may be used as a powerful tool to further investigate the role of KMP-11 during Leishmania development and/or as a diagnostic tool in Leishmania infection.

摘要

指数富集的配体系统进化(SELEX)程序是一种方法,其中单链寡核苷酸基于它们与靶分子的相互作用从各种各样的序列中被选择。我们设计了一种使用胶体金的新型SELEX方法,以选择针对婴儿利什曼原虫KMP-11的高亲和力单链DNA适配体。动质体膜蛋白-11(KMP-11)是动质体寄生虫细胞膜的主要成分。尽管其功能尚不清楚,但KMP-11是一种与细胞骨架相关的蛋白质这一事实表明它可能参与运动或鞭毛结构的其他方面。我们已经分离出了一个特异性结合婴儿利什曼原虫KMP-11的单链DNA适配体群体。该群体已在一系列体外实验中得到表征,表明它可能用作进一步研究KMP-11在利什曼原虫发育过程中的作用的有力工具和/或用作利什曼原虫感染的诊断工具。

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