رسالة جامعية
Expanding the molecular tools for the microalgae Chlorella vulgaris and Phaeodactylum tricornutum
| العنوان: | Expanding the molecular tools for the microalgae Chlorella vulgaris and Phaeodactylum tricornutum |
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| المؤلفون: | Hickland, Patrick |
| بيانات النشر: | University of Cambridge Department of Plant Sciences Darwin College |
| سنة النشر: | 2023 |
| المجموعة: | Apollo - University of Cambridge Repository |
| مصطلحات موضوعية: | microalgae |
| الوصف: | Many species of microalgae have attracted attention from industrial biotechnology with the goal of harnessing specific physiological traits in applications including fuel, food, feed and high value medicinal products. However, to realise their full potential certain species will require optimisation through engineering. This study focussed on molecular tool development to address bottlenecks hindering the engineering of two microalgal species: the green alga *Chlorella vulgaris* and the marine diatom *Phaeodactylum tricornutum*, both exhibit rapid growth rates and high density cultures making them prominent candidates for use in numerous applications. Work in this thesis was sponsored by the algal biotechnology company Algenuity, Stewartby, Bedfordshire. Progress in engineering of *Chlorella* has been slowed by the lack of reliable transformation protocols. In this study transformation protocols based on electroporation and biolistics were investigated with the goal of reliably transforming Algenuity’s proprietary *C. vulgaris* strain. The literature surrounding these transformation methods was systematically reviewed and informed the protocol development process. RNAseq analysis of the closely related species *Chlorella variabilis* informed the design and assembly of a Golden Gate (GG) based *Chlorella* MoClo toolkit including promoters, terminators and antibiotic resistance genes. Regulatory elements from a *Chlorella* virus and *Chlamydomonas reinhardtii* were also included in the toolkit. From these parts antibiotic resistance cassettes were assembled and used in trials to develop an electroporation or biolistic transformation protocol for *C. vulgaris*. Attempts to transform the proprietary strain by these means were unsuccessful but a transformation technique based on *Escherichia coli* conjugation was successfully employed enabling the characterisation of the parts assembled. Results of conjugative transformation suggest that use of the *C. reinhardtii* RbcS2 intron1 in antibiotic resistance CDS reduces ... |
| نوع الوثيقة: | doctoral or postdoctoral thesis |
| وصف الملف: | application/pdf |
| اللغة: | English |
| العلاقة: | https://www.repository.cam.ac.uk/handle/1810/362641Test; https://doi.org/10.17863/CAM.104724Test |
| DOI: | 10.17863/CAM.104724 |
| الإتاحة: | https://doi.org/10.17863/CAM.104724Test https://www.repository.cam.ac.uk/handle/1810/362641Test |
| حقوق: | All Rights Reserved ; https://www.rioxx.net/licenses/all-rights-reservedTest/ |
| رقم الانضمام: | edsbas.E9960D62 |
| قاعدة البيانات: | BASE |
| DOI: | 10.17863/CAM.104724 |
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