title: Transfection of Eimeria and Toxoplasma using heterologous regulatory sequences.
abstract: Eimeriatenella and Toxoplasmagondii 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.
authors.affiliation: Parasitology Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.forename: Xianyong
authors.initials: X
authors.lastname: Liu
authors.forename: Tuanyuan
authors.initials: T
authors.lastname: Shi
authors.forename: Xiaoxi
authors.initials: X
authors.lastname: Huang
authors.forename: Hongyan
authors.initials: H
authors.lastname: Wang
authors.forename: Lili
authors.initials: L
authors.lastname: Hao
authors.forename: Guangwen
authors.initials: G
authors.lastname: Yin
authors.forename: Xun
authors.initials: X
authors.lastname: Suo
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000378:metabolism; D002478:Cells, Cultured; D004539:Eimeria / Q000235:genetics* / Q000254:growth & development; D016786:Eimeria tenella / Q000235:genetics / Q000254:growth & development; D008018:Life Cycle Stages; D008164:Luminescent Proteins / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics; D050436:Regulatory Elements, Transcriptional / Q000235:genetics*; D014122:Toxoplasma / Q000235:genetics* / Q000254:growth & development; D014162:Transfection / Q000379:methods*; D014170:Transformation, Genetic
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria