Improving Pharmaceutical Protein Production in Oryza sativa

Kuo, Yung Che; Tan, Chia Chun; Ku, Jung Ting; Hsu, Wei Cho; Su, Sung Chieh; Lu, Chung An; Huang, Li

doi:10.3390/ijms14058719

Cited by 46 publications

(36 citation statements)

References 114 publications

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“…Transgenic technology can break reproductive isolation to realize gene transfer among different species, and rice is a potential plant that can be used as a recombinant protein expres- sion system (Liu et al, 2011;Kim et al, 2012;An et al, 2013;Kuo et al, 2013). However, after a high-expression gene in one species it is transferred to another species, the expression is often reduced greatly, particularly for species with a large expression difference, leading to the loss of the gene's function (Gustafsson et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

A novel synthetic Cry1Ab gene resists rice insect pests

Song

et al. 2014

Genet. Mol. Res.

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ABSTRACT. A few insect control genes of Bacillus thuringiensis have been modified successfully to increase the expression in plants by replacing rare codons, increasing GC content, and avoiding the DNA elements that could cause premature transcription termination, mRNA instability, and potential methylation. However, the modification process was intricate and often confused researchers. In this study, we adopted a simple method to modify Cry1Ab only by individually replacing its amino acid sequence with corresponding rice-preferred codons based on analysis of 92,188 coding DNA sequences. Unexpectedly, all elements of A+T richness, which terminate or destabilize transcription in plants, were avoided in the newly designed mCry1Ab. However, mCry1Ab had 2 notable features: less synonymous codons and high GC content. mCry1Ab only employed 22 of the 61 codons to encode protein and had an enhanced GC content of 65%. The increase in GC content caused abundant potential methylation signals to emerge in mCry1Ab. To test whether mCry1Ab could be expressed in rice, we transferred it into Oryza japonica variety A novel synthetic Cry1Ab gene Wanjing97. Insect bioassays revealed that transgenic plants harboring this gene driven by 2 promoters, CaMV35S and OsTSP I, were highly resistant to rice leaffolder (Cnaphalocrocis medinalis). Analysis of R 0 to R 2 generation plants indicated that the mCry1Ab was inherited stably by the progeny. Our study provided a simple modified method for expressing exogenous genes in rice and confirmed that less synonymous codons and high GC content do not affect transgene expression in rice.

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Section: Discussionmentioning

confidence: 99%

A novel synthetic Cry1Ab gene resists rice insect pests

Song

et al. 2014

Genet. Mol. Res.

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“…Verschiedene Pflanzen kommen als Expressionssysteme in Frage, darunter beispielsweise Erbsen (Mikschofsky/Broer, 2012), die Grünalge Chlamydomonas reinhardtii (RosalesMendoza et al, 2012), Reis (Kuo et al, 2013), verschiedene Nachtschattengewächse wie Kartoffel und Tomate (Rigano et al, 2013), oder Mais (Yusibov et al, 2011). Auch die Produktion von Vakzinen in pflanzlichen "hairy roots", die durch Transformation mit Agrobacterium rhizogenes gewonnen werden, wird diskutiert (Skarjinskaia et al, 2013) Kritik Agarwal et al, 2013).…”

Section: Plant Phenotypingunclassified

7. Themenbereich grüne Gentechnologie: Pflanzenzüchtung und Agrarwirtschaft

Müller‐Röber¹,

Marx-Stölting²

2015

Dritter Gentechnologiebericht

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“…형질전환 식물세포를 이용한 재조합 단백질 생산은 세포의 내부에 단백질을 축적해 생산하거나, 세포 외부로 단백질을 분비하여 생산한다 [9,10]. 세포 외부로의 재조합 단백질의 분비는 세포를 파쇄하지 않고 배지 내 단백질의 분석과 정제 를 할 수 있기 때문에 생물공정에 있어 유리하다 [11]. In situ 회수는 주로 미생물을 이용한 이차대사산물 생산 공정에서 생산량을 증진시키는데 이용되어 왔다.…”

Section: Introductionunclassified

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa

Yeol¹,

전수환²,

권준영³

et al. 2016

KSBB Journal

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In this research, recombinant human cytotoxic Tlymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

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Improving Pharmaceutical Protein Production in Oryza sativa

Cited by 46 publications

References 114 publications

A novel synthetic Cry1Ab gene resists rice insect pests

A novel synthetic Cry1Ab gene resists rice insect pests

7. Themenbereich grüne Gentechnologie: Pflanzenzüchtung und Agrarwirtschaft

In situ Recovery of hCTLA4Ig from Suspension Cell Cultures of Oryza sativa

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