无细胞蛋白质合成系统的研究进展

Advances in Cell-free Protein Biosynthesis

无细胞蛋白质合成系统是一种以外源mRNA或DNA为模板 ,通过在细胞抽提物的酶系中补充底物和能源物质来合成蛋白质的体外系统 .与传统的体内重组表达系统相比 ,体外无细胞合成系统具有多种优点 ,如可表达对细胞有毒害作用或含有非天然氨基酸 (如D 氨基酸 )的特殊蛋白质 ,能够直接以PCR产物作为模板同时平行合成多种蛋白质 ,开展高通量药物筛选和蛋白质组学的研究等 .本文综述了无细胞蛋白质合成系统的发展历史、系统中合成蛋白质所需的能量供应、遗传模板的稳定性和微型无细胞生物反应器等多方面的研究 ,并探讨了无细胞蛋白质合成系统中存在的难点。

The cell free protein synthesis system enables the direct in vitro synthesis of protein from template DNA or mRNA by using the existing enzymes in the cell extract, which is supplemented with substrates and energy sources. In comparison with the conventional recombinant cell technology, cell free protein synthesis system has several advantages, such as: the synthesis of some specific proteins with cytotoxicity or having several unnaturally modified amino acids ( D amino acids), simultaneous synthesis of many different protein molecules by using PCR generated linear DNA template, which satisfy the high throughput demand of proteomics and drug screening, etc. This review aims to retrieve the development history of cell free synthesis system, and to outline the research advances in energy regeneration, the stability of mRNA and the mini bioreactor for this system. Moreover, the present challenge, research directions and potential application prospects in this cell-free system will also be discussed.