학술논문

Risk assessment and bioburden evaluation of Agrobacterium tumefaciens-mediated transient protein expression in plants using the CaMV35S promoter
Document Type
article
Source
BMC Biotechnology, Vol 23, Iss 1, Pp 1-12 (2023)
Subject
Plant molecular farming
Process biosafety
Product toxicity
Promoter activity
Rhizobium radiobacter
Biotechnology
TP248.13-248.65
Language
English
ISSN
1472-6750
Abstract
Abstract Large-scale transient expression of recombinant proteins in plants is increasingly used and requires the multi-liter cultivation of Agrobacterium tumefaciens transformed with an expression vector, which is often cloned in Escherichia coli first. Depending on the promoter, unintentional activity can occur in both bacteria, which could pose a safety risk to the environment and operators if the protein is toxic. To assess the risk associated with transient expression, we first tested expression vectors containing the CaMV35S promoter known to be active in plants and bacteria, along with controls to measure the accumulation of the corresponding recombinant proteins. We found that, in both bacteria, even the stable model protein DsRed accumulated at levels near the detection limit of the sandwich ELISA (3.8 µg L−1). Higher levels were detected in short cultivations (