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Disruption of essential plastid gene expression caused by T7 RNA polymerase-mediated transcription of plastid transgenes during early seedling development

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Disruption of essential plastid gene expression caused by T7 RNA polymerase-mediated transcription of plastid transgenes during early seedling development

Content Provider	Springer Nature Link
Author	Magee, Alan M. MacLean, Daniel Gray, John C. Kavanagh, Tony A.
Copyright Year	2006
Abstract	Transcription of plastid transgenes by plastid-targeted T7 RNA polymerase (ptT7RNAP) during early seedling development in tobacco was associated with a pale-green leaf phenotype, depletion of plastid rRNAs and arrest of shoot development. Extensive analysis of mutant seedlings at the transcript level using DNA microarrays and RNA gel blotting revealed severe disruption of plastid rRNA accumulation at 4-days post-germination and reduced transcript accumulation for the essential gene clpP. Several nuclear genes encoding plastid proteins were differentially regulated in mutant seedlings over time. Ef-Tu was upregulated at 4-days post-germination and then subsequently downregulated, while RbcS was already downregulated at this early time point. The downregulation of nuclear genes encoding plastid proteins suggests disruption of plastid-to-nucleus signalling. In contrast, transcripts of three plastid genes showed increased accumulation in mutant seedlings. Transcripts of ndhC and ndhK accumulated at high levels possibly due to T7RNAP-mediated enhancement of transcription, while ptT7RNAP-mediated transcription through the phage T7 Tφ terminator into the adjacent plastome increased the level of accD transcripts. The leakiness of the Tφ terminator has implications for the use of T7RNAP-based expression systems in plastid biotechnology.
Starting Page	415
Ending Page	428
Page Count	14
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	16
Issue Number	4
e-ISSN	15739368
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2006-11-14
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Chloroplast Gene expression Microarray Plastid biotechnology Plastid transgene T7 RNA polymerase Human Genetics Plant Sciences Animal Genetics and Genomics Plant Genetics & Genomics Molecular Medicine
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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