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体外提高恶性疟原虫配子体的同步生产。

Improved synchronous production of Plasmodium falciparum gametocytes in vitro.

作者信息

Fivelman Quinton L, McRobert Louisa, Sharp Sarah, Taylor Cathy J, Saeed Maha, Swales Claire A, Sutherland Colin J, Baker David A

机构信息

Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, UK.

出版信息

Mol Biochem Parasitol. 2007 Jul;154(1):119-23. doi: 10.1016/j.molbiopara.2007.04.008. Epub 2007 Apr 20.

Abstract

The sexual stages of the Plasmodium falciparum life cycle are attractive targets for vaccines and transmission blocking drugs. Difficulties in culturing and obtaining large amounts of sexual stage P. falciparum parasites, particularly early stages, have often limited research progress in this area. We present a new protocol which simplifies the process of stimulating gametocytogenesis leading to improved synchronous gametocyte production. This new method can be adapted to enrich for early stage gametocytes (I and II) with a higher degree of purity than has previously been achieved, using MACS magnetic affinity columns. The protocol described lends itself to large scale culturing and harvesting of synchronous parasites suitable for biochemical assays, northern blots, flow cytometry, microarrays and proteomic analysis.

摘要

恶性疟原虫生命周期中的有性阶段是疫苗和传播阻断药物的诱人靶点。培养和获取大量有性阶段的恶性疟原虫寄生虫,尤其是早期阶段,存在困难,这常常限制了该领域的研究进展。我们提出了一种新方案,该方案简化了刺激配子体生成的过程,从而提高了同步配子体的产量。这种新方法可通过MACS磁亲和柱进行调整,以富集纯度更高的早期配子体(I期和II期),其纯度高于以往所达到的水平。所描述的方案适用于大规模培养和收获适合进行生化分析、Northern印迹、流式细胞术、微阵列和蛋白质组分析的同步寄生虫。

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