Efeito de candidatos a supressores de silenciamento gênico viral em expressão de proteína recombinate em plantas

AUTOR(ES)
DATA DE PUBLICAÇÃO

2010

RESUMO

The aim of this work was to increase the efficiency of transient heterolog protein expression system using viral gene-silencing suppressors and Agrobacterium integrated. The use of plants as bioreactor is a promising strategy for large scale production of the heterolog protein. However, the use of genetically modified plants may lead to low protein expression due to the strong system of degradation of the transgene mRNA, known as the Post-Transcriptional Gene Silencing (PTGS). PTGS is a defense mechanism against RNA viruses and retrotransposons in eukaryotic cells, however, this system is often counter-attacked by gene virus silencing suppressors during infections. To overcome the low level of expression of recombinant proteins in plants, the strategy of co-expression of the target protein with PTGS suppressors was considered. To evaluate the usefulness of this strategy, a reporter gene gfp, coding for the Green Fluorescent Protein (GFP) was cloned into binary vector for Agrobacterium tumefaciens transient expression (ATTE) and coinfiltred separately, with three PTGS suppressor candidates, 126K protein of Pepper mild mottle virus (Tobamovirus), Hc-Pro of Brugmansia suaveolans mottle virus (Potyvirus) and AC2 of Tomato severe rugose virus (Begomovirus). Our results indicate the feasibility of the strategy, demonstrating that co-expression of the target protein with a silencing supressor candidate leads to an increase on the heterologous protein expression in question.

ASSUNTO(S)

virologia biologia experimental ptgs supressores de silenciamento hc-pro 126k ac2 gfp genetica ptgs, gfp, silencing suppression, hc-pro, 126k, ac2

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