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将调节性DNA元素纳入病毒载体可以改善植物中重组蛋白质的表达

Ryan J Coates¹, Simon Scofield², Mark T Young³

¹School of Biosciences, Cardiff University, Sir Martin Evans Building, Museum Avenue, Cardiff, CF10 3AX, United Kingdom.

Scientific reports

|November 21, 2024

概括

暂时的植物表达系统可以改进以获得更高的产量. 使用基于TMV的pJL-TRBO载体,添加特定的5'非翻译区域 (UTR) 显著提高了重组蛋白质的产生.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
植物科学植物科学

背景情况:

植物为复杂,改性蛋白质提供可扩展和具有成本效益的重组蛋白质生产.
植物中的短暂表达速度更快,但与稳定的系统相比,蛋白质产量低,往往受到限制.

研究的目的:

通过优化基于TMV的pJL-TRBO载体,增强植物中的重组蛋白表达.
确定关键的遗传组件,有助于增加过渡植物表达系统中的蛋白质产量.

主要方法:

修改了pJL-TRBO向量,包括促进子,5' UTR,3' UTR,双终端子和矩阵附着区域等元素.
使用增强的绿色光蛋白 (eGFP) 作为记者来量化表达水平.
评估使用植物光成像,SDS-PAGE和西部涂抹的蛋白质产量.

主要成果:

优化的结构显示,与原始的pJL-TRBO-eGFP载体相比,复合蛋白表达的显著增加.
增强表达仅归因于添加了额外的5'未翻译区域 (UTR).
量化证实了通过光成像和生物化学测试的实质性产量改善.

结论:

添加特定的5'UTR是显著改善植物中短暂重组蛋白产量的关键因素.
开发的表达结构为高产量的植物性蛋白质生产提供了有价值的工具.
这项研究为生物技术应用植物表达载体设计的进一步进展提供了一个模板.

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