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Please use this identifier to cite or link to this item:
http://ir.lib.cyut.edu.tw:8080/handle/310901800/8368
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| Title: | Effectiveness and stability of heterologous proteins expressed in plants by Turnip mosaic virus vector at five different insertion sites |
| Authors: | 張清安
Chen, C. C.;Chen, T. C.;Raja, J. A. J.;Chang, C. A.;Chen, L. W.;Lin, S. S.;Yeh, S. D. |
| Contributors: | 生化科技研究所 |
| Keywords: | TuMV vector;Potyvirus;Plant expression;Dust mite allergen Der p 5;Insertion sites;Stability |
| Date: | 2007-12-31 |
| Issue Date: | 2010-12-02 09:43:17 (UTC+8) |
| Publisher: | Elsevier
http://www.sciencedirect.com/science/article/pii/S0168170207002286 |
| Abstract: | The N-terminal (NT) regions of particular protein-coding sequences are generally used for in-frame insertion of heterologous open reading
frames (ORFs) in potyviral vectors for protein expression in plants. An infectious cDNA clone of Turnip mosaic virus (TuMV) isolate YC5 was
engineered at the generally used NT regions of HC-Pro and CP, and other possibly permissive sites to investigate their effectiveness to express
the GFP (jellyfish green fluorescent protein) and Der p 5 (allergen from the dust mite, Dermatophagoides pteronyssinus) ORFs. The results
demonstrated the permissiveness of the NT regions of P3, CIP and NIb to carry the ORFs and express the translates as part of the viral polyprotein,
the processing of which released free-form proteins in the host cell milieu. However, these sites varied in their permissiveness to retain the ORFs
intact and hence affect the heterologous protein expression. Moreover, strong influence of the inserted ORF and host plants in determining the
permissiveness of a viral genomic context to stably carry the alien ORFs and hence to support their prolonged expression was also noticed. In
general, the engineered sites were relatively more permissive to the GFP ORF than to the Der p 5 ORF. Among the hosts, the local lesion host,
Chenopodium quinoa Willd. showed the highest extent of support to TuMV to stably carry the heterologous ORFs at the engineered sites and the
protein expression therefrom. Among the systemic hosts, Nicotiana benthamiana Domin proved more supportive to TuMV to carry and express
the heterologous ORFs than the Brassica hosts, whereas the protein expression levels were significantly higher and more stable in the plants of
Brassica campestris L. var. chinensis and B. campestris L. var. ching-geeng than those in the plants of B. juncea L. and B. campestris L. var.
pekinensis. |
| Relation: | Virus Research 130, p.210-227 |
| Appears in Collections: | [應用化學系] 期刊/會議/專書論文
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