Tackling heterogeneity: a leaf disc-based assay for the high-throughput screening of transient gene expression in tobacco.

ABSTRACT: Transient Agrobacterium-mediated gene expression assays for Nicotiana tabacum (N. tabacum) are frequently used because they facilitate the comparison of multiple expression constructs regarding their capacity for maximum recombinant protein production. However, for three model proteins, we found that recombinant protein accumulation (rpa) was significantly influenced by leaf age and leaf position effects. The ratio between the highest and lowest amount of protein accumulation (max/min ratio) was found to be as high as 11. Therefore, construct-based impacts on the rpa level that are less than 11-fold will be masked by background noise. To address this problem, we developed a leaf disc-based screening assay and infiltration device that allows the rpa level in a whole tobacco plant to be reli

SUBMITTER: Piotrzkowski N 

PROVIDER: S-EPMC3448687 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature