請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/46196
標題: Effectiveness and stability of heterologous proteins expressed in plants by Turnip mosaic virus vector at five different insertion sites
作者: Chen, C.C.
葉錫東
Chen, T.C.
Raja, J.A.
Chang, C.A.
Chen, L.W.
Lin, S.S.
Yeh, S.D.
關鍵字: TuMV vector
potyvirus
plant expression
dust mite allergen Der p 5
insertion sites
stability
helper component-proteinase
tobacco etch potyvirus
long-distance
movement
to-cell movement
capsid protein
beta-glucuronidase
molecular-biology
escherichia-coli
foreign proteins
raphanus-sativus
期刊/報告no:: Virus Research, Volume 130, Issue 1-2, Page(s) 210-227.
摘要: 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. (c) 2007 Elsevier B.V. All rights reserved.
URI: http://hdl.handle.net/11455/46196
ISSN: 0168-1702
文章連結: http://dx.doi.org/10.1016/j.virusres.2007.06.014
顯示於類別:植物病理學系

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