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Novel tools for plastid engineering in higher plants
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Title
Novel tools for plastid engineering in higher plants
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Yu
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Qiguo
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1989-
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Qiguo Yu
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Pal Maliga
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Dong
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Juan
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Juan Dong
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Gallavotti
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Andrea
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Andrea Gallavotti
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Nickels
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Bryce
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Rutgers University
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School of Graduate Studies
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theses
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ETD doctoral
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2020
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2020-01
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2020
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English
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Plastid genome engineering holds great promise in many biotechnological applications, including production of vaccines and antibodies, conferring insect tolerance by expressing dsRNAs, implanting novel metabolic pathways and improving photosynthetic efficiency and so on. However, the plastid transformation technology is not yet available in most crop plants.
I was part of the group that developed a system for plastid transformation in Arabidopsis thaliana, a Brassicaceae species, that is potentially applicable to the important oilseed crop canola (Brassica napus) and vegetable brassicas such as broccoli, cauliflower and cabbage (Brassica oleracea). Efficient plastid transformation is enabled by eliminating a duplicate fatty acid biosynthetic pathway in chloroplasts.
High level expression of recombinant proteins is desirable, but it may cripple the plant growth. To achieve on demand expression of plastid transgenes, I was involved in a project that exploited the use of an engineered PPR (pentatricopeptide) RNA-binding protein for post-transcriptional regulation of chloroplast genes. One application is ethanol-inducible expression of chloroplast proteins by activating mRNA translation when the protein is needed. Thus, the transplastomic plants can be grown full size in the absence of the inducer and expression is turned on during the protein production phase. The second application is tuber-specific expression of plastid genes where plastid gene expression is normally very law. When the engineered PPR variant is expressed from a tuber-specific patatin promoter, a 60-fold increase over 0.02%, the maximum protein yield achieved to date, has been obtained in the potato tuber.
Plastids represent an appealing target for synthetic biology applications, where predictable protein output is of paramount importance. I designed synthetic operons from which reporter gene expression could be obtained in tobacco chloroplasts over a large dynamic range by post-transcriptional regulation, using the right combination of RNA binding proteins, RNA-binding protein binding sites and RNA processing elements.
I was part of the group that developed a system for plastid transformation in Arabidopsis thaliana, a Brassicaceae species, that is potentially applicable to the important oilseed crop canola (Brassica napus) and vegetable brassicas such as broccoli, cauliflower and cabbage (Brassica oleracea). Efficient plastid transformation is enabled by eliminating a duplicate fatty acid biosynthetic pathway in chloroplasts.
High level expression of recombinant proteins is desirable, but it may cripple the plant growth. To achieve on demand expression of plastid transgenes, I was involved in a project that exploited the use of an engineered PPR (pentatricopeptide) RNA-binding protein for post-transcriptional regulation of chloroplast genes. One application is ethanol-inducible expression of chloroplast proteins by activating mRNA translation when the protein is needed. Thus, the transplastomic plants can be grown full size in the absence of the inducer and expression is turned on during the protein production phase. The second application is tuber-specific expression of plastid genes where plastid gene expression is normally very law. When the engineered PPR variant is expressed from a tuber-specific patatin promoter, a 60-fold increase over 0.02%, the maximum protein yield achieved to date, has been obtained in the potato tuber.
Plastids represent an appealing target for synthetic biology applications, where predictable protein output is of paramount importance. I designed synthetic operons from which reporter gene expression could be obtained in tobacco chloroplasts over a large dynamic range by post-transcriptional regulation, using the right combination of RNA binding proteins, RNA-binding protein binding sites and RNA processing elements.
Subject
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Topic
Plant Biology
Subject
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Topic
Chloroplast engineering higher plants
Subject
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Topic
Plastids -- Genetic engineering
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Rutgers University Electronic Theses and Dissertations
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School of Graduate Studies Electronic Theses and Dissertations
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rucore10001600001
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doi:10.7282/t3-3gcq-ba29
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156 pages : illustrations
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Ph.D.
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Includes bibliographical references
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The author owns the copyright to this work.
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Yu
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Qiguo
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2019-12-17 12:04:14
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Qiguo Yu
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Rutgers University. School of Graduate Studies
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Author Agreement License
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
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2020-01-31
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2021-01-30
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Access to this PDF has been restricted at the author's request. It will be publicly available after January 30th, 2021.
Copyright
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Copyright protected
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Open
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Permission or license
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