Cloning and characterization of the cDNA encoding the venom serine protease found in Polistes dominulus (Old world paper wasp) / by Christina D. Fitch.
| Author/creator | Fitch, Christina D. author. |
| Other author | Hoffman, Donald R., degree supervisor. |
| Other author | Schmidt, Margit, degree supervisor. |
| Other author | East Carolina University. Department of Biology. |
| Format | Theses and dissertations |
| Production | 2003. |
| Description | 76 leaves : illustrations (some color) ; 28 cm |
| Supplemental Content | Access via ScholarShip |
| Subjects |
| Summary | Hymenoptera, a medically important order of insects, are responsible for the death of approximately forty individuals per year in the United States. It has been calculated that upward of 0.4% of the population has an insect allergy and this allergy is usually to more than one insect. Polistes dominulus (Old World Paper Wasp), a paper wasp indigenous to the Mediterranean region, has begun to establish itself in the North Eastern part of the United States and has become one of the dominant species. Three major allergens have been identified from the venom of paper wasps, phospholipase AiB, hyaluronidase and Antigen 5. The isolation of the serine protease found in Polistes dominulus and Polistes exclamans (New World Paper Wasp) was accomplished through affinity chromatography, and allowed for the study of IgE binding from patients who had been sensitized by the old world species and the new world species. These assays suggested that while the venom protease from the old world species may be a major allergenic component, the protease from the new world appears to be a weaker allergen. Using the partial amino acid sequence obtained from protein sequencing, analysis was done using reverse transcriptase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The data were analyzed and the resulting sequences were overlayed to construct a full-length sequence. The constructed sequence was used to gain a full-length clone that was transformed into a eukaryotic (Pichia pastoris) expression vector. Expression was not successful, there were a number of variables that may have contributed to this. The sequence that was transformed lacked a propeptide region, without this sequence the protein is active and may degrade itself. Another factor maybe a possible internal Poly A site found in the coding sequence of the serine protease, which may lead to early termination of the transcript, producing a protein that can not be purified. The acquisition of the cDNA sequence will aid in the procuring the complete structure of this molecule, an investigation of allergenicity and comparison of the sequence to that of other venom serine proteases. |
| General note | Presented to the faculty of the Department of Biology. |
| General note | Advisor: Donald R. Hoffman |
| General note | Advisor: Margit Schmidt |
| Dissertation note | M.S. East Carolina University 2003 |
| Bibliography note | Includes bibliographical references (leaves 74-76). |
| Genre/form | dissertations. |
| Genre/form | Academic theses. |
| Genre/form | Academic theses. |
| Genre/form | Thèses et écrits académiques. |