title: Babesia bovis: gene isolation and characterization using a mung bean nuclease-derived expression library.
abstract: Genomic DNA prepared from erythrocyte cultures of Babesia bovis merozoites was digested with mung bean nuclease and used to construct a lambda gt11 expression library of B. bovis recombinants. Immunoscreening with two polyclonal antibody probes detected multiple recombinants from which two, designated Bb-1 and Bb-3, were chosen for further analysis. Monospecific immunoglobulins isolated from the screening sera using nitrocellulose-bound fusion proteins were employed to determine the native molecular weight and the intracellular location of the babesial proteins encoded by the recombinants. Clone Bb-1 encodes an antigen of 77,000 Da located at the apical end of the intraerythrocytic parasite. A protein of 75,000 Da encoded by clone Bb-3 is associated with the infected red blood cell cytoplasm and/or membrane but not with the merozoite.
authors.affiliation: Department of Veterinary Microbiology and Parasitology, Texas A&M University, College Station 77843.
authors.forename: C A
authors.initials: CA
authors.lastname: Tripp
authors.forename: G G
authors.initials: GG
authors.lastname: Wagner
authors.forename: A C
authors.initials: AC
authors.lastname: Rice-Ficht
mesh_terms: D000818:Animals; D001403:Babesia / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D015723:Gene Library; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification; D015183:Restriction Mapping; D015719:Single-Strand Specific DNA and RNA Endonucleases
chemical_list: D011506:Proteins; D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases