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Expression of a Functional Recombinant Human Basic Fibroblast Growth Factor from Transgenic Rice Seeds

机译:转基因水稻种子中功能重组人碱性成纤维细胞生长因子的表达

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摘要

Basic fibroblast growth factor (FGF-2) is an important member of the FGF gene family. It is widely used in clinical applications for scald and wound healing in order to stimulate cell proliferation. Further it is applied for inhibiting stem cell differentiation in cultures. Due to a shortage of plasma and low expression levels of recombinant rbFGF in conventional gene expression systems, we explored the production of recombinant rbFGF in rice grains (Oryza sativa bFGF, OsrbFGF). An expression level of up to 185.66 mg/kg in brown rice was obtained. A simple purification protocol was established with final recovery of 4.49% and resulting in a yield of OsrbFGF reaching up to 8.33 mg/kg OsrbFGF. The functional assay of OsrbFGF indicated that the stimulating cell proliferation activity on NIH/3T3 was the same as with commercialized rbFGF. Wound healing in vivo of OsrbFGF is equivalent to commercialized rbFGF. Our results indicate that rice endosperm is capable of expressing small molecular mass proteins, such as bFGF. This again demonstrates that rice endosperm is a promising system to express various biopharmaceutical proteins.
机译:碱性成纤维细胞生长因子(FGF-2)是FGF基因家族的重要成员。它被广泛用于烫伤和伤口愈合的临床应用中,以刺激细胞增殖。此外,它被用于抑制培养物中的干细胞分化。由于血浆缺乏和常规基因表达系统中重组rbFGF的低表达水平,我们探索了水稻籽粒(Oryza sativa bFGF,OsrbFGF)中重组rbFGF的生产。在糙米中的表达水平高达185.66 mg / kg。建立了简单的纯化方案,最终回收率为4.49%,导致OsrbFGF的产量高达8.33 mg / kg OsrbFGF。 OsrbFGF的功能测定表明,刺激NIH / 3T3的细胞增殖活性与商业化rbFGF相同。 OsrbFGF在体内的伤口愈合相当于商业化的rbFGF。我们的结果表明,稻胚乳能够表达小分子蛋白质,例如bFGF。这再次证明稻胚乳是表达各种生物药物蛋白的有前途的系统。

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