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Published on: February 20, 2015
Differential display analysis of gene expression in two immunologically distinct strains of Eimeria maxima
1Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, N1G 2W1, Ontario, Canada, mfernand@uoguelph.ca.
Abstract:
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.
Insights
Researchers identified a unique gene fragment, GS-453, expressed in Eimeria maxima Guelph strain (GS) but not in the M6 strain. This discovery aids in understanding genetic differences between distinct Eimeria maxima strains.
Area of Science:
- Molecular Biology
- Parasitology
- Genomics
Background:
- Eimeria maxima strains exhibit immunological distinctness.
- Understanding genetic variations is crucial for parasite control.
Purpose of the Study:
- To identify differentially expressed genes between two Eimeria maxima strains (GS and M6).
- To characterize a novel gene fragment unique to the GS strain.
Main Methods:
- mRNA differential display technique was employed.
- Complementary DNA (cDNA) cloning and sequencing of the GS-453 fragment.
- Northern and Southern blot analyses were performed.
- Bioinformatic database searches were conducted.
Main Results:
- A 453-bp cDNA fragment (GS-453) was isolated, present exclusively in GS sporozoites.
- The corresponding mRNA (approx. 1.6 kb) was detected in GS but absent in M6.
- Genomic Southern blots revealed sequence similarities but distinct hybridization patterns between strains.
- No homology was found with existing sequences in public databases.
Conclusions:
- The GS-453 fragment represents a strain-specific genetic marker in Eimeria maxima.
- This finding contributes to the molecular characterization of distinct Eimeria maxima strains.
- The novel sequence highlights potential targets for strain differentiation and control strategies.
