Transient expression of DNA and RNA in parasitic helminths by using particle bombardment

被引:86
作者
Davis, RE [1 ]
Parra, A
LoVerde, PT
Ribeiro, E
Glorioso, G
Hodgson, S
机构
[1] Fordham Univ, Dept Biol Sci, Bronx, NY 10458 USA
[2] SUNY Buffalo, Dept Microbiol, Buffalo, NY 14214 USA
关键词
transfection; Ascaris; schistosome; splice leader RNA; firefly luciferase;
D O I
10.1073/pnas.96.15.8687
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Parasitic helminths (worms belonging to several metazoan phyla) cause considerable morbidity and mortality in humans. They are an important veterinary problem, and they result in significant economic losses in animal grazing and agriculture. Experimental studies on parasitic helminths have been limited by a lack of parasite cell lines and methods for molecular genetic analyses. We evaluated particle bombardment (biolistics) as a strategy to introduce and express nucleic acids in these multicellular parasites. By using embryos of the parasitic nematode Ascaris as a model, we developed methods to introduce and express both DNA and RNA during several stages of Ascaris embryogenesis, Biolistic transfection will facilitate experimental strategies in Ascaris embryos complementing other biochemical tools available (e.g., in vitro whole-cell embryo extracts for transcription, RNA processing, and translation), Transfection experiments with adult schistosomes further suggest that the biolistic strategy should be applicable to a variety of other parasitic helminths, The development of these methods provides molecular genetic tools to study gene expression and the biology of a variety of types and developmental stages of important helminth parasites.
引用
收藏
页码:8687 / 8692
页数:6
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