Simple and reliable system for transient gene expression for the characteristic signal sequences and the estimation of the localization of target protein in plant cellстатья

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[1] Simple and reliable system for transient gene expression for the characteristic signal sequences and the estimation of the localization of target protein in plant cell / A. A. Tyurin, K. V. Kabardaeva, M. A. Berestovoy et al. // Russian Journal of Plant Physiology. — 2017. — Vol. 64, no. 5. — P. 672–679. The efficiency of transient gene expression in plants credibly demonstrated characteristics of gene functions in numerous studies. Two key strategies of transient expression became favorites among research- ers: protoplast transfection and agroinfiltration. Each of them, alongside the advantages, has its own con- straints. In this work, an easy, rapid, and reliable system for characterization of the signal sequences and determinations of target protein localization in a plant cell is proposed and tested. This system—called the AgI–PrI—implies production of protoplasts from plant tissues after agroinfiltration. Reliability of the pro- posed system for transient gene expression has been proved using characterized signal sequences in Nicotiana benthamiana cells. The corresponding protocol is less expensive and depends to a lesser degree on the profes- sional skills in the area of protoplast isolation and transfection; furthermore, it may be applicable to other plant species with either available efficient methods of agroinfiltration and protoplast isolation or with the potential for one of the protocols to be supplemented. Thus, the AgI–PrI technique makes it possible to com- bine the advantages of two widely used methods for the transient gene expression in plants—agroinfiltration and protoplast isolation and transfection—and concurrently avoids their critical points. [ DOI ]

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