Babesia bovis clones: biochemical and enzymatic characterization
Studies were undertaken to generate additional knowledge of the biochemistry of Babesia bovis. A modified in vitro culture technique used for cloning B. bovis. This technique included a low oxygen concentration atmosphere (2%, O/sub 2/, 5% CO/sub 2/, 93% N/sub 2/) and 4 mm fluid level. Cultures initiated with one infected erythrocyte were maintained until parasitemias of positive wells reached 2% parasitemia. Primary clones were obtained and from these, nine clones were recloned twice and used for subsequent studies. A procedure was developed to concentrate and separate B. bovis merozoites and infected erythrocytes by Percoll density gradients. Merozoites separated at 1.087 g/ml specific density, whereas infected erythrocytes separated at 1.121 g/ml. Viability of purified parasites was not affected. Agarose gel electrophoresis was used to identify metabolic enzyme in B. bovis and B. bigemina. The enzymes LDH, GDH, GPI and HK were detected in both species. Molecular analysis by one and two-dimensional gel electrophoresis of proteins metabolically labeled with /sup 35/S-methionine indicated that two clones, derived from the same field strain, were similar but not identical to the parent. Fewer proteins were observed in the parental strain. Growth of two 60-Co irradiated B. bovis clones indicated a dose-effect relationship. Growth of parasites exposed for the longest period was initially retarded but returned to normal growth after two or three subcultures. Cultures exposed for shorter periods were unaffected with respect to the rate of growth. Analysis of electrophoretic mobility of metabolic enzyme showed a change in migration pattern.
- Research Organization:
- Missouri Univ., Columbia (USA)
- OSTI ID:
- 6516563
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
Similar Records
Inhibitor-bound complexes of dihydrofolate reductase-thymidylate synthase from Babesia bovis
A newly characterized malaria antigen on erythrocyte and merozoite surfaces induces parasite inhibitory antibodies
Related Subjects
59 BASIC BIOLOGICAL SCIENCES
BABESIDAE
BIOCHEMISTRY
BIOLOGICAL RADIATION EFFECTS
HEMIACETAL DEHYDROGENASES
ELECTROPHORESIS
CLONING
COBALT 60
DOSE-RESPONSE RELATIONSHIPS
ENZYME ACTIVITY
ERYTHROCYTES
GAMMA RADIATION
LACTATE DEHYDROGENASE
METHIONINE
OXYGEN
PARASITES
SULFUR 35
AMINO ACIDS
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOLOGICAL MATERIALS
BLOOD
BLOOD CELLS
BODY FLUIDS
CARBOXYLIC ACIDS
CHEMISTRY
COBALT ISOTOPES
DAYS LIVING RADIOISOTOPES
DRUGS
ELECTROMAGNETIC RADIATION
ELEMENTS
ENZYMES
EVEN-ODD NUCLEI
INTERMEDIATE MASS NUCLEI
INTERNAL CONVERSION RADIOISOTOPES
INVERTEBRATES
IONIZING RADIATIONS
ISOMERIC TRANSITION ISOTOPES
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
MATERIALS
MICROORGANISMS
MINUTES LIVING RADIOISOTOPES
NONMETALS
NUCLEI
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
OXIDOREDUCTASES
PROTOZOA
RADIATION EFFECTS
RADIATIONS
RADIOISOTOPES
SPOROZOA
SULFUR ISOTOPES
YEARS LIVING RADIOISOTOPES
560130* - Radiation Effects on Microorganisms
550201 - Biochemistry- Tracer Techniques