Transient expression of beta-galactosidase in differentiating sporozoites of Eimeria tenella.
A transient transfection system has been developed for a member of the Apicomplexa, Eimeria tenella, using beta-galactosidase (betagal) from Escherichia coli as the reporter enzyme. Successfully expressed constructs contained sequences of the E. tenella microneme gene Etmic-1 fused to the coding region of lacZ. Transfectants expressing betagal were able to invade host cells and proceed through part of the life-cycle, forming schizonts from which merozoites were released. This indicated that transfectants could differentiate at least to first generation schizonts. However, this differentiation was delayed compared with unelectroporated sporozoites by approximately 15 h. Some merozoites arising from transfected sporozoites also expressed betagal. These results are encouraging for the development of a stable transfection system for E. tenella, using betagal as a reporter enzyme.
Kelleher
M
M
Division of Molecular Biology, The Institute for Animal Health, Compton, Berkshire, UK.
Tomley
F M
FM
D000818:Animals; D019677:Artificial Gene Fusion / Q000379:methods; D001483:Base Sequence; D002460:Cell Line / Q000469:parasitology; D016786:Eimeria tenella / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D018274:Electroporation; D017930:Genes, Reporter / Q000235:genetics; D008018:Life Cycle Stages / Q000502:physiology; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013997:Time Factors; D014162:Transfection / Q000379:methods; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism*
C117852:Etmic-1 protein, Eimeria tenella; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D001616:beta-Galactosidase