Immunological and molecular characterization of conserved nuclear RNA binding proteins in Physarum polycephalum /
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| Other Authors: | , |
| Format: | Thesis Book |
| Language: | English |
| Published: |
1989.
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| Subjects: | |
| Online Access: | Link to OAKTrust copy |
| Abstract: | The 34 Kd Physarum polycephalum nucleolar protein B-36 was first identified because of its similarities to the hnRNP core proteins. These similarities include a high glycine content and the presence of the rare amino acid asymmetric dimethylarginine. Its localization to the nucleolus has been confirmed using monoclonal antibodies (Christensen et al., 1986). This and other data implicate it in rRNA packaging and processing. In an effort to identify a B-36 cDNA clone, a Physarum Agtll cDNA expression library was screened with the polyclonal antiserum 792, and several potential clones were identified. The clones were rescreened and those that remained positive were plaque purified. The four clones that gave the strongest positive reactions were then subjected to an antibody selection assay, since polyclonal 792 cross-reacts with other Physarum proteins. The polyclonal 792 antibodies were eluted from nitrocellulose filter plaque lifts containing individual clones, and were then used to screen Western transfers of Physarum nuclear proteins. In three cases the eluted antibodies were apparently specific for B-36. One of these clones (lb) was chosen for further analysis. Restriction enzyme mapping indicated that clone lb had two AccI sites that divided the clone into three pieces of approximately equal size. Sequencing of lb indicated that the clone contained a large repeat but showed no homology to the B-36 protein sequence already available. The clone did have one region that coded for a peptide region with a high content of arginine and glycine. It is this region that is suspected to cross-react with an epitope in B-36 that is recognized by the polyclonal antibody 792. Several families of oligonucleotide primers were synthesized based on the available amino acid sequence for B-36.. |
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| Item Description: | Typescript (photocopy). Vita. "Major subject: Biology." |
| Physical Description: | x, 111 leaves : illustrations ; 29 cm |
| Bibliography: | Includes bibliographical references. |