DNA probes detect genomic diversity in Theileria parva stocks

• Published in: Molecular and biochemical parasitology Vol. 25; no. 3; pp. 213 - 226
• Main Authors: Conrad, Patricia A., Iams, Keith, Brown, Wendy C., Sohanpal, Baljinder, ole-MoiYoi, Onesmo K.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier B.V 01.10.1987; Elsevier Science
• Subjects: Animals; Apicomplexa - genetics; Biological and medical sciences; calves; Cattle; DNA; DNA - analysis; DNA - genetics; DNA probes; DNA Restriction Enzymes; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; Genes; genomics; Hybridization; Nucleic Acid Hybridization; OFAGE; Parasite characterization; Polymorphism, Genetic; Polymorphism, Restriction Fragment Length; Protozoa; Restriction fragment length polymorphism; Systematics. Geographical distribution. Morphology. Cytology; Theileria parva; Theileriasis - parasitology; stb; PBS; cpm; SSC; SDS; DNA probes; Theileria parva; ECF; TNE; Hybridization; EDTA; Tris; TE; OFAGE; Restriction fragment length polymorphism; TBE; kb; Parasite characterization; Protozoa; Gel electrophoresis; Restriction fragment; Identification; DNA; Blotting assay; Molecular probe; Southern blotting; Pathogen; Sporozoa; Stock; Polymorphism
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/0166-6851(87)90085-5

Different stocks of Theileria parva were analysed for restriction fragment length polymorphisms by agarose gel electrophoresis, orthogonal-field-alternation gel electrophoresis (OFAGE) and Southern hybridization with DNA probes. Polymorphisms seen with DNA from purified piroplasms of different T. parva stocks, after digestion with restriction enzymes, were more clearly apparent with OFAGE than with standard agarose gel electrophoresis. Genomic differences between these theilerial parasites were investigated further using three DNA probes, which were selected from a genomic library of T. parva (Muguga) piroplasm DNA cloned in λgt11. All three clones hybridized to T. parva DNA in preparations from schizont-infected bovine lymphoblastoid cells and to DNA from intraerythrocytic piroplasms. These probes did not, however, hybridize under high stringency conditions to DNA prepared from uninfected bovine lymphoblasts, T. mutans piroplasms, or bovine lymphoblasts infected with T. annulata or T. taurotragi. The five Kenyan stocks of T. parva that were tested showed characteristic hybridization patterns with these DNA probes. Our results show that DNA probes can be used to distinguish selected stocks of T. parva by hybridization to DNA either from intraerythrocytic piroplasms taken from infected cattle, or from isolates of schizont-infected bovine lymphoblastoid cells that are maintained continuously in vitro.