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Elastin-like polypeptide and gamma-zein fusions significantly increase recombinant protein accumulation in soybean seeds

文献类型： 外文期刊

作者： Yang, Jing 1 ; Xun, HongWei 1 ; Niu, Lu 1 ; He, Hongli 2 ; Cheng, Yunqing 2 ; Zhong, Xiaofang 1 ; Zhao, Qianqian 1 ; Xing 1 ;

作者机构： 1.Jilin Acad Agr Sci, Jilin Prov Key Lab Agr Biotechnol, Changchun 130033, Peoples R China

2.Jilin Normal Univ, Siping 136000, Peoples R China

关键词： Soybean seed; Fusion partner; Elastin-like polypeptide; Γ -Zein; Recombinant protein

期刊名称：TRANSGENIC RESEARCH （ 影响因子：1.856； 五年影响因子：1.974 ）

ISSN： 0962-8819

年卷期：

页码：

收录情况： SCI

摘要： Soybean seeds are an ideal host for the production of recombinant proteins because of their high content of proteins, long-term stability of seed proteins under ambient conditions, and easy establishment of efficient purification protocols. In this study, a polypeptide fusion strategy was applied to explore the capacity of elastin-like polypeptide (ELP) and gamma-zein fusions in increasing the accumulation of the recombinant protein in soybean seeds. Transgenic soybean plants were generated to express the gamma-zein- or ELP-fused green fluorescent protein (GFP) under the control of the soybean seed-specific promoter of beta-conglycinin alpha subunit (BCSP). Significant differences were observed in the accumulation of zein-GFP and GFP-ELP from that of the unfused GFP in transgenic soybean seeds based on the total soluble protein (TSP), despite the low-copy of T-DNA insertions and similar expression at the mRNA levels in selected transgenic lines. The average levels of zein-GFP and GFP-ELP accumulated in immature seeds of these transgenic lines were 0.99% and 0.29% TSP, respectively, compared with 0.07% TSP of the unfused GFP. In mature soybean seeds, the accumulation of zein-GFP and GFP-ELP proteins was 1.8% and 0.84% TSP, an increase of 3.91- and 1.82-fold, respectively, in comparison with that of the unfused GFP (0.46% TSP). Confocal laser scanning analysis showed that both zein-GFP and GFP-ELP were abundantly deposited in many small spherical particles of transgenic seeds, while there were fewer such florescence signals in the same cellular compartments of the unfused GFP-expressing seeds. Despite increased recombinant protein accumulation, there were no significant changes in the total protein and oil content in seeds between the transgenic and non-transformed plants, suggesting the possible presence of threshold limits of total protein accumulation in transgenic soybean seeds. Overall, our results indicate that gamma-zein and ELP fusions significantly increased the accumulation of the recombinant protein, but exhibited no significant influence on the total protein and oil content in soybean seeds.

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