A1 Refereed original research article in a scientific journal

HandyGun: An improved custom-designed, non-vacuum gene gun suitable for virus inoculation




AuthorsSikorskaite S, Vuorinen AL, Rajamaki ML, Nieminen A, Gaba V, Valkonen JPT

PublisherELSEVIER SCIENCE BV

Publication year2010

Journal: Journal of Virological Methods

Journal name in sourceJOURNAL OF VIROLOGICAL METHODS

Journal acronymJ VIROL METHODS

Volume165

Issue2

First page 320

Last page324

Number of pages5

ISSN0166-0934

DOIhttps://doi.org/10.1016/j.jviromet.2010.02.025


Abstract

Particle bombardment with a non-vacuum gene gun is an efficient method for transfection of plant cells with cloned viruses and initiation of virus infection. The HandyGun developed in this study is an improved version of a non-vacuum gene gun. Bombardment parameters were studied by inoculating an infectious, 355 promoter-driven cDNA of Potato virus A (PVA; Potyvirus) to the potato clone 'A6'. Nicotiana benthamiana and N. tabacum as plasmid DNA coated on microprojectiles (gold particles). The large number of initial infection sites (necrotic local lesions) observed on inoculated 'A6' leaves and the high percentage of Nicotiana plants which were infected systemically with PVA following inoculation with HandyGun were not particularly sensitive to variation in the parameters tested (helium pressure and the amounts of plasmid DNA and gold particles). Data showed that HandyGun is a robust and reliable tool for obtaining high infection rates in plants reproducibly. It is easy and inexpensive to use and can be constructed from parts commonly available. (C) 2010 Elsevier B.V. All rights reserved.



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