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Amplicon-plus targeting technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants
BibTeX
@MISC{Weissinger_amplicon-plustargeting,
author = {Arthur K. Weissinger},
title = {Amplicon-plus targeting technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants},
year = {}
}
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Abstract
Abstract High-level expression of transgenes is esse-ntial for cost-effective production of valuable phar-maceutical proteins in plants. However, transgenic proteins often accumulate in plants at low levels. Low levels of protein accumulation can be caused by many factors including post-transcriptional gene silencing (PTGS) and/or rapid turnover of the transgenic pro-teins. We have developed an Amplicon-plus Targeting Technology (APTT), by using novel combination of known techniques that appears to overcome both of these factors. By using this technology, we have suc-cessfully expressed the highly-labile L1 protein of canine oral papillomavirus (COPV L1) by infecting transgenic tobacco plants expressing a suppressor of post-transcriptional gene silencing (PTGS) with a PVX amplicon carrying a gene encoding L1, and targeting the vaccine protein into the chloroplasts. Further, a scalable ‘‘wound-and-agrospray’ ’ inoculation method has been developed that will permit high-throughput Agrobacterium inoculation of Nicotiana tabacum, and a spray-only method (named ‘‘agrospray’’) for use with N. benthamiana to allow large-scale application of this technology. The good yield and short interval from inoculation to harvest characteristic of APTT, com-bined with the potential for high-throughput achieved by use of the agrospray inoculation protocol, make this system a very promising technology for producing high value recombinant proteins, especially those known to be highly labile, in plants for a wide range of applications including producing vaccines against rapidly evolving pathogens and for the rapid response needed to meet bio-defense emergencies.
Keyphrases
unstable vaccine protein rapid production tobacco plant low level post-transcriptional gene silencing spray-only method protein accumulation rapid turnover transgenic pro-teins novel combination transgenic protein high value recombinant protein wide range cost-effective production valuable phar-maceutical protein vaccine protein many factor short interval abstract high-level expression canine oral papillomavirus agrospray inoculation protocol scalable wound-and-agrospray inoculation method high-throughput agrobacterium inoculation pvx amplicon large-scale application promising technology copv l1 nicotiana tabacum bio-defense emergency highly-labile l1 protein transgenic tobacco plant rapid response good yield amplicon-plus targeting technology
