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首页> 外文期刊>BMC Biotechnology >Protein body formation in stable transgenic tobacco expressing elastin-like polypeptide and hydrophobin fusion proteins
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Protein body formation in stable transgenic tobacco expressing elastin-like polypeptide and hydrophobin fusion proteins

机译：表达弹性蛋白样多肽和疏水蛋白融合蛋白的稳定转基因烟草中蛋白质的形成

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Background Plants are recognized as an efficient and inexpensive system to produce valuable recombinant proteins. Two different strategies have been commonly used for the expression of recombinant proteins in plants: transient expression mediated by Agrobacterium ; or stable transformation of the plant genome. However, the use of plants as bioreactors still faces two main limitations: low accumulation levels of some recombinant proteins and lack of efficient purification methods. Elastin-like polypeptide (ELP), hydrophobin I (HFBI) and Zera? are three fusion partners found to increase the accumulation levels of recombinant proteins and induce the formation of protein bodies (PBs) in leaves when targeted to the endoplasmic reticulum (ER) in transient expression assays. In this study the effects of ELP and HFBI fusion tags on recombinant protein accumulation levels and PB formation was examined in stable transgenic Nicotiana tabacum . Results The accumulation of recombinant protein and PB formation was evaluated in two cultivars of Nicotiana tabacum transformed with green fluorescent protein (GFP) fused to ELP or HFBI, both targeted and retrieved to the ER. The ELP and HFBI tags increased the accumulation of the recombinant protein and induced the formation of PBs in leaves of stable transgenic plants from both cultivars. Furthermore, these tags induced the formation of PBs in a concentration-dependent manner, where a specific level of recombinant protein accumulation was required for PBs to appear. Moreover, agro-infiltration of plants accumulating low levels of recombinant protein with p19, a suppressor of post-transcriptional gene silencing ( PTGS ), increased accumulation levels in four independent transgenic lines, suggesting that PTGS might have caused the low accumulation levels in these plants. Conclusion The use of ELP and HFBI tags as fusion partners in stable transgenic plants of tobacco is feasible and promising. In a constitutive environment, these tags increase the accumulation levels of the recombinant protein and induce the formation of PBs regardless of the cultivar used. However, a specific level of recombinant protein accumulation needs to be reached for PBs to form.
机译：背景技术植物被认为是生产有价值的重组蛋白的有效且廉价的系统。两种不同的策略通常用于在植物中表达重组蛋白：农杆菌介导的瞬时表达;或植物基因组的稳定转化。然而，将植物用作生物反应器仍然面临两个主要限制：一些重组蛋白的低积累水平和缺乏有效的纯化方法。弹性蛋白样多肽（ELP），疏水蛋白I（HFBI）和Zera？是三个融合伙伴，在瞬时表达测定中靶向内质网（ER）时，发现它们会增加重组蛋白的积累水平并诱导叶片中蛋白质体（PBs）的形成。在这项研究中，在稳定的转基因烟草中研究了ELP和HFBI融合标签对重组蛋白积累水平和PB形成的影响。结果评价了两个转化为绿色荧光蛋白（GFP）的烟草（Nicotiana tabacum）的重组蛋白的积累和PB的形成。 ELP和HFBI标签增加了重组蛋白的积累并诱导了两个品种稳定转基因植物叶片中PB的形成。此外，这些标签以浓度依赖性的方式诱导PB的形成，其中PB的出现需要特定水平的重组蛋白积累。此外，p19是转录后基因沉默（PTGS）的抑制剂，对农业中的重组蛋白水平低的植物进行浸润，增加了四个独立转基因品系的积累水平，这表明PTGS可能导致了这些植物中的低积累水平。 。结论在稳定的烟草转基因植物中使用ELP和HFBI标签作为融合伴侣是可行且有前途的。在组成性环境中，无论使用哪种品种，这些标签都会增加重组蛋白的积累水平并诱导PBs的形成。但是，要形成PB，需要达到特定水平的重组蛋白积累。

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