Transfection of Schistosoma mansoni by electroporation and the description of a new promoter sequence for transgene expression

被引:31
作者
Correnti, Jason M. [1 ]
Jung, Euihye [1 ]
Freitas, Tori C. [1 ]
Pearce, Edward J. [1 ]
机构
[1] Univ Penn, Sch Vet Med, Dept Pathobiol, Philadelphia, PA 19104 USA
关键词
helminth parasite; transfection; promoter; terminator; electroporation;
D O I
10.1016/j.ijpara.2007.02.011
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
We sought to investigate the efficacy of electroporation for the introduction of plasmid-based DNA constructs into Schistosoma mansoni, and expanded our study to examine parameters governing transgene expression, including requirements of a 5' and 3' flanking sequence, as well as parasite developmental effects on transgene expression. We used luciferase as a reporter gene for this application. Our data show that electroporation allows the transfection of immature schistosomes, and defines 5' promoter sequence from the schistosome actin gene (SmAct1.1), coupled promiscuously with various 3' terminator sequences, as a powerful promoter of transgene expression in growing, but not early non-growing, schistosomula. The methodology described herein will facilitate ectopic expression of genes of interest in schistosomes. (c) 2007 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:1107 / 1115
页数:9
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