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A Viral Suppressor P1/HC-Pro Increases the GFP Gene Expression in Agrobacterium-mediated Transient Assay

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Abstract

More than 20 post-transcriptional gene silencing (PTGS) suppressors have been found since HC-Pro, the first gene silencing suppressor, was found in 1998. The silencing suppressor strongly suggested that gene silencing functions as natural defense mechanisms against viruses. It also represented a valuable tool for the dissection of the gene silencing pathway. We have used P1/HC-Pro RNA silencing suppressor activity to increase green fluorescent protein (GFP) expression in tobacco using an Agrobacterium-mediated transient expression system. P1/HC-Pro stimulated GFP-gene expression but not dsGFP-gene expression was shown by RT-PCR, Northern and Western blot analysis. Expression of the gene silencing suppressor and the target gene provided a new strategy of heterogeneous gene expressing in plants. It may be of commercial significance to produce foreign proteins using plant bioreactors.

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Abbreviations

HC-Pro:

helper component-proteinase

PTGS:

post-transcriptional gene silencing

RISC:

RNA-induced silencing complex

RT-PCR:

reverse transcription-PCR

siRNAs:

short-interfering RNAs

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Acknowledgement

This work was supported by the Program for Changjiang Scholars and Innovative Research Team (PCSIRT) in University (#IRT0519), the Programme of Introducing Talents of Discipline to Universities 111 Project B07017 and the Chinese National Plant Transformation Center (JY03-B-17). We thank Dr. Carrington, Institute of Biological Chemistry, Washington State University, for providing pSLJ75515-smGFP, pSLJ75515-dsGFP and pSLJ75515-0027 constructs; Dr. Baulcombe, the Sainsbury Lab, UK, for N. benthamiana.

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Authors and Affiliations

  1. Institute of Genetics and Cytology, Northeast Normal University, Changchun, 130024, China

    Pengda Ma, Jinying Liu, Hongxia He, Meiying Yang, Meina Li, Xiaojuan Zhu & Xingzhi Wang

  2. Biotechnology Research Center, Jilin Academy of Agricultural Sciences, Changchun, 130033, China

    Pengda Ma

  3. School of Life Sciences, Jilin Agricultural University, Changchun, 130118, China

    Meiying Yang

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  1. Pengda Ma
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Correspondence to Xingzhi Wang.

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Ma, P., Liu, J., He, H. et al. A Viral Suppressor P1/HC-Pro Increases the GFP Gene Expression in Agrobacterium-mediated Transient Assay. Appl Biochem Biotechnol 158, 243–252 (2009). https://doi.org/10.1007/s12010-008-8332-y

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