Inducible expression of human papillomavirus‐16 L1 capsomeres in the plastomes of Nicotiana tabacum : Transplastomic plants develop normal flowers and pollen
| Autor: | Neelam Batool, Andreas G. Lössl, Sara Latif, Muhammad Sameeullah, Kehkshan Gull, Johanna Gottschamel, Martin Müller, Tahira Syed, Bushra Mirza, Mohammad Tahir Waheed, Filipe Mariz, Iqra Younus |
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| Rok vydání: | 2021 |
| Předmět: |
Male
0106 biological sciences Signal peptide Nicotiana tabacum Transgene Biomedical Engineering Bioengineering Flowers 01 natural sciences Applied Microbiology and Biotechnology 03 medical and health sciences 010608 biotechnology Tobacco Drug Discovery Humans Gene 030304 developmental biology Southern blot Human papillomavirus 16 0303 health sciences Ethanol biology Process Chemistry and Technology fungi food and beverages Oncogene Proteins Viral General Medicine Plants Genetically Modified biology.organism_classification Molecular biology Chloroplast Transformation (genetics) Pollen Molecular Medicine Capsid Proteins Female Biotechnology Transplastomic plant |
| Zdroj: | Biotechnology and Applied Biochemistry. 69:596-611 |
| ISSN: | 1470-8744 0885-4513 |
| Popis: | Human papillomavirus type-16 (HPV-16) is the major HPV type involved in causing cervical cancer among women. The disease burden is high in developing and underdeveloped countries. Previously, the constitutive expression of HPV-16 L1 protein led to male sterility in transplastomic tobacco plants. Here, the HPV-16 L1 gene was expressed in chloroplasts of Nicotiana tabacum under the control of an ethanol-inducible promoter, trans-activated by nucleus-derived signal peptide. Plants containing nuclear component were transformed with transformation vector pEXP-T7-L1 by biolistic gun. The transformation and homoplasmic status of transformed plants was verified by polymerase chain reaction and Southern blotting, respectively. Protein was induced by spraying 5% ethanol for 7 consecutive days. The correct folding of L1 protein was confirmed by antigen-capture ELISA using a conformation-specific antibody. The L1 protein accumulated up to 3 μg/g of fresh plant material. The L1 protein was further purified using affinity chromatography. All transplastomic plants developed normal flowers and produced viable seeds upon self-pollination. Pollens also showed completely normal structure under light microscope and scanning electron microscopy. These data confirm the use of the inducible expression as plant-safe approach for expressing transgenes in plants, especially those genes that cause detrimental effects on plant growth and morphology. |
| Databáze: | OpenAIRE |
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