Transfection of Eimeria and Toxoplasma using heterologous regulatory sequences
Transfection of Eimeria and Toxoplasma using heterologous regulatory sequences
复制标题
使用异源调控序列转染艾美球虫和弓形虫。
DOI：
10.1016/j.ijpara.2009.03.006
复制
发表时间：
2009-09-01
影响因子：
4
通讯作者：
Suo, Xun
中科院分区：
文献类型：
--
作者：
Zou, Jun;Liu, Xianyong;Suo, Xun
柔嫩艾美耳球虫和刚地弓形虫是顶复门原虫，在生物学和基因组学上有许多相似之处。虽然后一种寄生虫很容易在体外培养和遗传操作，但许多艾美耳球虫物种很难在体外生长。我们假设T.弓形虫可用于艾美耳球虫的研究。在这里，我们表明，三个不同的启动子序列来源于E。tenella不仅在艾美耳球虫属的其他物种（组蛋白H4）中有效地发挥作用，而且在T. gondii（组蛋白H4、肌动蛋白和微管蛋白）。同样，弓形虫的“管家”基因（微管蛋白）和差异调节基因（表面抗原基因，sag1）的启动子在E. tenella。外源调控序列的转染效率与同源启动子相似，但外源启动子的启动子强度略弱于同源启动子。tenella和弓形虫。结果表明，5'端调控序列不仅在艾美耳球虫种间，而且在弓形虫和艾美耳球虫之间都是功能保守的。tenella和T. gondii可作为一种新的艾美耳球虫根载体的转染检测系统，促进艾美耳球虫反向遗传学的发展。(C)2009年澳大利亚寄生虫学会由爱思唯尔有限公司出版。保留所有权利。
Eimeria tenella and Toxoplasma gondii are Apicomplexan protozoa and share many similarities in biology and genomics. While the latter parasites are easily cultured in vitro and genetically manipulated, many Eimeria species are difficult to grow in vitro. We hypothesised that molecular tools for the genetic manipulation of T. gondii could be applied to the study of Eimeria parasites. Here we show that three different promoter sequences originating from E. tenella could function effectively not only in other species of the Eimeria genus (histone H4) but also in T. gondii (histone H4, actin and tubulin). Similarly, promoters of the "housekeeping" gene (tubulin) and differentially regulated gene (surface antigen gene, sag1) of T gondii were effective in driving the expression of the yellow fluorescent protein (YFP) maker gene in E. tenella. The transfection efficiency with heterologous regulatory sequences was similar to that with homologous promoters; while the promoter strength of heterologous vectors is slightly weaker than the homologous vectors in both E. tenella and T gondii. The results suggest that 5' regulatory sequences are functionally conserved not only among the Eimeria species, but also between T gondii and E. tenella, and that T. gondii could be used as a novel transfection check system for Eimeria-rooted vectors, accelerating the development of reverse genetics in Eimeria spp. (C) 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.