Differential display analysis of gene expression in two immunologically distinct strains of Eimeria maxima.
Gene expression during sporulation and sporozoite excystation of two strains of Eimeria maxima was analyzed using the mRNA differential display technique. The two strains, the Guelph strain (GS) and a single sporocyst-derived strain (M6) from Florida, have been shown to be immunologically distinct. We isolated and cloned a 453-bp complimentary DNA (cDNA) fragment (GS-453) found only in GS sporozoites. In GS, this mRNA begins to be expressed during the earliest stages of oocyst sporulation and is continuously expressed up to and including in the excysted sporozoite. In all Northern blots, digoxigenin (DIG)-labeled GS-453 probe recognized an mRNA of approximately 1.6 kb from GS but not from RNA of M6. Southern blots using various endonucleases and probed with DIG-labeled GS-453 demonstrated that the genomes of both strains contained sufficiently similar sequences to permit hybridization with the probe, but the pattern of hybridization differed between the two strains. Extensive searches of the GenBank, European Molecular Biology Laboratory, and various apicomplexan expressed sequence tag databases using the DNA or inferred amino acid sequences of GS-453 cDNA clone did not identify similarity to any existing sequences.
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, N1G 2W1, Ontario, Canada, mfernand@uoguelph.ca.
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SC
Basak
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Lee
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JR
Barta
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Fernando
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002645:Chickens; D003048:Coccidiosis / Q000469:parasitology / Q000662:veterinary*; D016054:DNA, Protozoan / Q000235:genetics; D004539:Eimeria / Q000235:genetics* / Q000502:physiology; D020869:Gene Expression Profiling; D008969:Molecular Sequence Data; D011201:Poultry Diseases / Q000469:parasitology*; D012333:RNA, Messenger / Q000032:analysis; D016053:RNA, Protozoan / Q000032:analysis; D013045:Species Specificity; D033761:Spores, Protozoan / Q000235:genetics; D034101:Sporozoites / Q000378:metabolism; D013997:Time Factors
D016054:DNA, Protozoan; D012333:RNA, Messenger; D016053:RNA, Protozoan