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Stability of Potato virus X expression vectors is related to insert size: implications for replications models and risk assesment

Avesani, Linda and Marconi, Gianpiero and Morandini, Francesca and Albertini, Emidio and Bruschetta, Matteo and Bortesi, Luisa and Pezzotti, Mario and Porceddu, Andrea (2007) Stability of Potato virus X expression vectors is related to insert size: implications for replications models and risk assesment. Transgenic Research, Vol. 16 (5), p. 578-587. eISSN 1573-9368. Article.

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DOI: 10.1007/s11248-006-9051-1

Abstract

We investigated the stability of expression constructs based on Potato virus X (PVX) as a function of insert length. Five different inserts ranging in length from 261 to 1,758 bp (human proinsulin, murine interleukin-10, HIV-1 nef, petunia expansin-1 and human gad65) were expressed using a PVX vector in Nicotiana benthamiana plants for three sequential passages. Using a competitive RT-PCR approach we demonstrated that insert–deletion could occur in the first infection cycle for all inserts, but that this was much more likely to be the case for longer ones. This suggested a negative correlation between insert length and vector stability. Sequence analysis of the deleted constructs suggested that recombination usually occurred at sites close to the duplicated sub-genomic promoter, but in a smaller number of cases the foreign gene itself was probably involved, resulting in partially deleted constructs containing transgene fragments. The implications of these results in the context of recombinant protein expression and its risks are discussed.

Item Type:Article
ID Code:4140
Status:Published
Refereed:Yes
Uncontrolled Keywords:Potato virus X, transient expression, recombinant proteins, viral replication, risk assessment
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/07 Genetica agraria
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze agronomiche e genetica vegetale agraria
Publisher:Springer
eISSN:1573-9368
Copyright Holders:© Springer Science+Business Media B.V. 2006
Deposited On:09 Jul 2010 13:55

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