Agrobacterium-Mediated Transient Expression as an Approach to Production of Recombinant Proteins in Plants

Sheludko, Y. V.

doi:10.2174/187220808786241033

Cited by 47 publications

(36 citation statements)

References 88 publications

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“…In fact, a transgenic plant-based production of a foreign protein is low and often less than 1% of TSP (Sheludko, 2008). To overcome this low productivity, the chloroplast transformation technology was introduced and constantly progressed.…”

Section: Resultsmentioning

confidence: 99%

Effect of Rice stripe virus NS3 on Transient Gene Expression and Transgene Co-Silencing

Sohn¹,

Huh²,

Kim³

et al. 2011

The Plant Pathology Journal

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Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.

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Section: Resultsmentioning

confidence: 99%

Effect of Rice stripe virus NS3 on Transient Gene Expression and Transgene Co-Silencing

Sohn¹,

Huh²,

Kim³

et al. 2011

The Plant Pathology Journal

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“…Salah satu metode transformasi tanaman yang banyak digunakan adalah dengan bantuan Agrobacterium tumefaciens yang memiliki kemampuan secara biologis dalam mentransfer bagian DNA tertentu (T-DNA) dari plasmid Ti (tumor inducing) ke dalam sel tanaman yang kemudian berintegrasi ke genom tanaman target (Sheludko, 2008). Penggunaan A. tumefaciens sebagai vektor untuk melakukan transformasi genetik pada berbagai tanaman dikotil dan monokotil telah terbukti efektif dan mempunyai tingkat keberhasilan dan kestabilan gen yang tinggi, spesifik, mengurangi terbentuknya tanaman kimera dan lebih ekonomis dibandingkan metode transformasi genetik yang lain (Kumar et al, 2014;Mano et al, 2014;Sugiharto & Safitri, 2011).…”

Section: Pendahuluanunclassified

Evaluasi varietas, sumber eksplan dan strain Agrobacterium terhadap keberhasilan transformasi tebu dengan gen P5CS Evaluation of varieties, explant sources, and Agrobacterium strains for successful sugarcane transformation using P5CS gene

MINARSIH¹,

SUBIYARTI²,

RIYADI³

et al. 2016

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Genetic transformation can be used as an alter-native to develop sugarcane (Saccharum officinarum L.) tolerant to drought stress. P5CS gene has a role in biosynthesis of proline, an amino acid that accumulated under drought stress conditions. Transfer of a P5CS gene construct into plant cells in conjunction with regeneration of transgenic plantlets may develop sugarcane tolerant to drought stress. The aim of this research was to obtain an optimal transformation method which includes a suitable strain of Agrobacterium tumefaciens, and the best sugarcane explant and variety. The results showed that transfer of P5CS gene has been successfully carried out on sugarcane explants from solid media-derived calli, embryogenic calli and somatic embryos derived from temporary immersion system (TIS) culture. Whilst Agrobacterium strain LBA4404 was indicated as the most effective transformation vector. The regeneration of Kidang Kencana variety transformants from calli and somatic embryos was better than those of PS 881 and PS 891. The best performance of transformants based on the source of explants obtained from somatic embryos from TIS culture. Moreover, a succesfull Agrobacterium mediated transformation on sugarcane was indicated by transient expression of Gus gene and the ability of the transformants grew in a selection medium containing 50 ppm of kanamycin.Abstrak Transformasi genetik dapat digunakan sebagai upaya untuk merakit tebu (Saccharum officinarum L.) toleran terhadap cekaman kekeringan. Gen P5CS diketahui berperan dalam biosintesis prolin, yaitu asam amino yang umumnya terakumulasi ketika tanaman mengalami cekaman kekeringan. Transfor-masi gen P5CS dan regenerasi transgeniknya mungkin dapat menghasilkan tanaman tebu trans-genik yang toleran terhadap cekaman kekeringan. Tujuan penelitian ini adalah untuk mendapatkan metode transformasi yang optimum yang mencakup strain Agrobacterium tumefaciens yang sesuai, sumber eksplan dan varietas tebu terbaik sebagai target transformasi. Hasil penelitian menunjukkan bahwa transformasi gen P5CS telah berhasil dilakukan ke eksplan tebu baik yang berupa kalus asal media padat maupun kalus embriogenik dan embrio somatik asal kultur sistem perendaman sesaat (SPS). Sementara itu strain A. tumefaciens LBA4404 menunjukkan hasil yang paling efektif sebagai vektor transformasi. Pertumbuhan transforman baik pada kalus maupun embrio somatik pada varietas Kidang Kencana terlihat paling baik dibandingkan dengan varietas PS 881 dan PS 891. Sumber eksplan yang paling efektif adalah embrio somatik yang diperoleh dari kultur SPS. Keberhasilan transformasi tebu me-lalui Agrobacterium ditunjukkan oleh ekspresi transien dari gen GUS dan kemampuan dari trans-forman untuk tumbuh di media yang mengandung 50 ppm kanamisin.

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“…Then, it became attractive for producing biopharmaceuticals due to their high protein yields and to not generate a GMO (Circelli et al 2010 ;Sheludko 2008 ). Another advantage of this system is that it allows the rapid co-expression of multiple genes (Huang and Mason 2004 ;Medrano et al 2009 ).…”

Section: Transient Expressionmentioning

confidence: 99%

Molecular Farming in Plants

Álvarez

2014

Plant Biotechnology for Health

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Molecular farming can generally be defi ned as the production of molecules (proteins, fatty acids) for the pharmaceutical and chemical industries in transgenic organisms (plants, animals, etc.).Molecular farming in plants has advantageous aspects as biosecurity, they do not bear pathogens for humans or animals, and they do not produce toxins. Also, plant protein synthetic machinery is able to produce complex glycosylated proteins as they have a glycosylation pattern with slight difference respect of that of mammals, and can perform foldings.When displayed in in vitro conditions, molecular farming have the characteristic advantages of that type of culture, mainly the capacity of working under Good Manufactory practices as is required by the pharmaceutical industry. The critical aspects of plants for molecular farming are the different glycosylation patterns respect to mammals, the duration of the productive process, and the relatively low yields.The approval by the FDA of the fi rst medicine to be used in humans, taliglucerase alfa (ELELYSO®), has give an impulse to this technology but there are some drawbacks to be addressed, particularly related to low yields and regulatory aspects.

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Agrobacterium-Mediated Transient Expression as an Approach to Production of Recombinant Proteins in Plants

Cited by 47 publications

References 88 publications

Effect of Rice stripe virus NS3 on Transient Gene Expression and Transgene Co-Silencing

Effect of Rice stripe virus NS3 on Transient Gene Expression and Transgene Co-Silencing

Evaluasi varietas, sumber eksplan dan strain Agrobacterium terhadap keberhasilan transformasi tebu dengan gen P5CS Evaluation of varieties, explant sources, and Agrobacterium strains for successful sugarcane transformation using P5CS gene

Molecular Farming in Plants

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