Towards A new molecular tool for transgenic plants: Control of protein biosynthesis by a seed-specific promoter
首发时间:2013-01-10
Abstract:[Background] Plants have been proven to be promising bioreactors in the production of many heterologous proteins including those in the biopharmaceutical industry. However, the efficiency of a potential plant bioreactor is impaired by the limitation in available promoters to drive foreign gene expression. Hence, the discovery of novel promoters is urgently needed. Seeds are ideal for protein storage and proteins can be kept as a stable form for several months. Thus a strong seed-specific promoter is a powerful tool in directing the expression and accumulation of target proteins in the seed. [Results] A 784-bp 5'-flanking sequence of the mung bean seed storage protein 8S globulin α' subunit gene was isolated by genome walking. When the sequence was analyzed with bioinformatics tools, numerous putative cis-elements were identified. The green fluorescent protein (GFP) reporter driven from this promoter was transiently expressed in protoplasts derived from mung bean cotyledons. Finally, transgenic Arabidopsis plants expressing the β-glucuronidase (GUS) reporter gene driven from the 8S globulin alpha' promoter were generated. GUS expression in transgenic seeds was observed by histochemical and quantitative GUS assays, confirming seed expression and high transcriptional activity. This study has identified the mung bean 8S α' promoter having potential for directing expression in seeds for bioreactor application.
keywords: Seed promoter genome walking mung bean.
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转基因植物的新分子工具:采用种子特异启动子调控蛋白合成
摘要:[背景]植物已经被证明是有前途的生物反应器,用于包括生物制药行业的许多外源蛋白的生产。然而,一个潜在的植物生物反应器的效率受限于启动子驱动外源基因表达的效率。因此,迫切需要发现新的启动子资源。种子是理想的蛋白存储场所,蛋白可保持数个月以上的活性。因此,一个高表达活性的种子特异性启动子可成为强大的工具,调控种子中的蛋白的表达和积累。[结果] 采用基因组布移的方法,分离得到绿豆种子贮藏蛋白8S球蛋白α'亚基基因的784 bp的5'-侧翼序列。序列分析和生物信息学分析表明,其包含了众多的顺式调控元件。该启动子可以成功驱动绿色荧光蛋白(GFP)在绿豆子叶原生质体的瞬时表达。该启动子驱动的β-葡萄糖醛酸苷酶(GUS)报告基因的表达也在转基因拟南芥中得到确认。GUS基因表达的转基因种子通过GUS组织化学和定量分析,表明了种子的特异表达及较高的转录活性。本研究表明绿豆8Sα'启动子具有可用于种子生物反应器的应用前景。
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