title: Cloning and partial characterization of the Cr32 gene of Cowdria ruminantium.
abstract: Cowdria organisms were purified by density gradient centrifugation. The DNA was used to construct expression libraries. The immunodominant Cr32 protein was purified and its N-terminal amino acid sequence was determined. The expression libraries were screened with Cr32-specific monoclonal antibodies, but did not yield Cr32-positive clones. Therefore a part of the Cr32-gene was amplified using primers derived from the N-terminal and an internal amino acid sequence. This DNA was used as a probe to detect the genomic DNA fragment encoding the Cr32 protein. This fragment was cloned, using genomic DNA of the Senegal strain of Cowdria ruminantium. A part of the gene comprising two third of its total length has been expressed in vector pGEX2T. This expression product is recognized by Cr32-specific monoclonal antibodies.
authors.affiliation: Department of Bacteriology, Faculty of Veterinary Medicine, University of Utrecht, Pays-Bas.
authors.forename: A H
authors.initials: AH
authors.lastname: van Vliet
authors.forename: F
authors.initials: F
authors.lastname: Jongejan
authors.forename: M
authors.initials: M
authors.lastname: van Kleef
authors.forename: B A
authors.initials: BA
authors.lastname: van der Zeijst
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D016995:Ehrlichia ruminantium / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D016056:Immunodominant Epitopes / Q000235:genetics
chemical_list: D016056:Immunodominant Epitopes