Degree Name
Master of Science (MS)
Semester of Degree Completion
1987
Thesis Director
Russell Carlson
Abstract
Serogroup specific antigens of Bradyrhizobium japonicum 61A123 were isolated using an immunochemical assay. Bacterial antigens were isolated by phenol-water extraction. These antigens were detected by inhibition of an enzyme linked immunosorbant assay (ELISA). Low pressure chromatography and digestion with proteinase K served to purify the antigens. Chemical composition was determined by gas chromatography (GC), mass spectrometry (MS), and colorimetric assays. The antigens were also characterized by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, and immunological properties determined by ELISA. These antigens were shown to be lipopolysaccharides (LPSs) containing fucose, a 2-amino-2,6-dideoxy hexose (unidentified), glucose and mannose as the major sugars, along with small amounts (<1%) of 2-keto-3-deoxyoctonate (KDO). Fatty acid determination showed the presence of 3-hydroxy myristic, lauric, oleic, and two unidentified fatty acids as major components. The growth medium for the bacteria was modified to supress the excessive production of extracellular polysaccharides (EPSs). This medium had no qualitative effect on the nodulation or immunochemical characteristics of the bacteria. Five hours of mild acid hydrolysis (1% acetic acid at 100°C) was required to separate the lipid from the polysaccharide portion of the molecule.
Recommended Citation
Carrion, Miguel Eduardo, "Isolation and Partial Characterization of Serogroup Specific Antigens from Bradyrhizobium japonicum 61A123" (1987). Masters Theses. 2594.
https://thekeep.eiu.edu/theses/2594
