Expression of Histophilus somni IbpA DR2 protective antigen in the diatom Thalassiosira pseudonana
| Autor: | Lynette B. Corbeil, Aubrey K. Davis, Mark Hildebrand, Lauren T. Crum |
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| Rok vydání: | 2017 |
| Předmět: |
0106 biological sciences
0301 basic medicine Silicon Light Cell division Thalassiosira pseudonana Cattle Diseases 01 natural sciences Applied Microbiology and Biotechnology Microbiology Cell wall Mice 03 medical and health sciences Immune system Antigen Transcription (biology) Protein purification Microalgae Animals Biotechnologically Relevant Enzymes and Proteins IbpA DR2 Respiratory Tract Infections Diatoms Antigens Bacterial Vaccines Synthetic biology Diatom General Medicine Carbon Dioxide Histophilus somni biology.organism_classification Antibodies Bacterial Recombinant Proteins 030104 developmental biology Bacterial Vaccines Cattle Pasteurellaceae Pasteurellaceae Infections Valuable proteins Transcription Factors Protective antigen 010606 plant biology & botany Biotechnology |
| Zdroj: | Applied Microbiology and Biotechnology |
| ISSN: | 1432-0614 0175-7598 |
| Popis: | Increasing demand for the low-cost production of valuable proteins has stimulated development of novel expression systems. Many challenges faced by existing technology may be overcome by using unicellular microalgae as an expression platform due to their ability to be cultivated rapidly, inexpensively, and in large scale. Diatoms are a particularly productive type of unicellular algae showing promise as production organisms. Here, we report the development of an expression system in the diatom Thalassiosira pseudonana by expressing the protective IbpA DR2 antigen from Histophilus somni for the production of a vaccine against bovine respiratory disease. The utilization of diatoms with their typically silicified cell walls permitted development of silicon-responsive transcription elements to induce protein expression. Specifically, we demonstrate that transcription elements from the silicon transporter gene SIT1 are sufficient to drive high levels of IbpA DR2 expression during silicon limitation and growth arrest. These culture conditions eliminate the flux of cellular resources into cell division processes, yet do not limit protein expression. In addition to improving protein expression levels by molecular manipulations, yield was dramatically increased through cultivation enhancement including elevated light and CO2 supplementation. We substantially increased recombinant protein production over starting levels to 1.2% of the total sodium dodecyl sulfate-extractable protein in T. pseudonana, which was sufficient to conduct preliminary immunization trials in mice. Mice exposed to 5 μg of diatom-expressed DR2 in whole or sonicated cells (without protein purification) exhibited a modest immune response without the addition of adjuvant. Electronic supplementary material The online version of this article (doi:10.1007/s00253-017-8267-8) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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