Transfection of Eimeria and Toxoplasma using heterologous regulatory sequences.
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.
Zou
Jun
J
Parasitology Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Liu
Xianyong
X
Shi
Tuanyuan
T
Huang
Xiaoxi
X
Wang
Hongyan
H
Hao
Lili
L
Yin
Guangwen
G
Suo
Xun
X
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
D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria