title: Co-expression of reporter genes in the widespread pathogen Eimeria tenella using a double-cassette expression vector strategy.
abstract: The double-cassette expression vector strategy is valuable for many studies, including comparative analysis of the function of promoters and expression of genes in different compartments. In this study, we report co-expression of enhanced yellow fluorescent protein (EYFP) and red fluorescent protein (RFP) in Eimeria tenella transfected with two double-cassette expression vectors, pMIC-EYFP/ACT-RFP and pMIC-EYFP/ACTss-RFP. The results showed that under regulation of the mic1 promoter, EYFP was expressed in sporulated oocysts but not in unsporulated ones, while under regulation of the actin promoter RFP was expressed in both forms. We found that the signal peptide of Toxoplasma gondii dense granule protein 8 (GRA8) located the RFP expression to the parasitophorous vacuoles of the parasites, the margins of the unsporulated oocysts and the cavities of the sporocysts. The feasibility of co-expression of exogenous proteins in E. tenella is important for the development of transgenic E. tenella as a novel vaccine vector.
authors.affiliation: National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
authors.forename: Guangwen
authors.initials: G
authors.lastname: Yin
authors.forename: Xianyong
authors.initials: X
authors.lastname: Liu
authors.forename: Jun
authors.initials: J
authors.lastname: Zou
authors.forename: Xiaoxi
authors.initials: X
authors.lastname: Huang
authors.forename: Xun
authors.initials: X
authors.lastname: Suo
mesh_terms: D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016786:Eimeria tenella / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D011401:Promoter Regions, Genetic; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C054562:yellow fluorescent protein, Bacteria