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United States Department of Agriculture

Agricultural Research Service

Title: Molecular Cloning and Baculovirus Expression of a 19 Kilodalton Eimeria Protein Conserved on Sporozoites and Merozoites of Several Eimeria Species.

item Lillehoj, Hyun
item Choi, Kang
item Jenkins, Mark
item Vakharia, V - UNIV OF MARYLAND

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: May 1, 1998
Publication Date: N/A

Technical Abstract: A cDNA encoding a 19kDa protein of Eimeria which is present on both sporozoites and merozoites has been cloned and expressed in E. coli and baculovirus expression system to investigate its immunogenicity. Clone 3-1E contained a cDNA insert of 1086 bp and a poly A tail. E. coli containing a recombinant pBluescript 3-1E plasmid and SF9 cells containing a 3-1E plasmid were grown and cell lysates prepared. A 19-21 kDa protein was identified in a cell lysate derived from E. coli harboring a 3-1E plasmid, and a 22-24 kDa protein was recognized in a SF9 cell lysate containing a 3-1E plasmid in Western blots. Western blot analysis also revealed that this protein is present on both sporozoites and merozoites of E. acervulina. IFA staining showed positive staining with sporozoites of several Eimeria species. Spleen lymphocytes and serum from chickens which had been hyperimmunized with E. acervulina showed an antigen-specific T cell and B cell responses to 3-1E encoded protein respectively. Immunization of chickens with recombinant coccidia protein from SF9 cells transfected with 3-1E plasmid induced a substantial reduction in oocyst production. These results indicate that 3-1E encodes a highly conserved Eimeria protein which is immunogenic.

Last Modified: 4/18/2015