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固相无细胞蛋白质合成及其在生物技术中的应用。

Solid-Phase Cell-Free Protein Synthesis and Its Applications in Biotechnology.

机构信息

Heterogeneous Biocatalysis Laboratory, Center for Cooperative Research in Biomaterials (CIC biomaGUNE), Basque Research and Technology Alliance (BRTA), Donostia-San Sebastián, Spain.

出版信息

Adv Biochem Eng Biotechnol. 2023;185:21-46. doi: 10.1007/10_2023_226.

Abstract

Cell-free systems for the in vitro production of proteins have revolutionized the synthetic biology field. In the last decade, this technology is gaining momentum in molecular biology, biotechnology, biomedicine and even education. Materials science has burst into the field of in vitro protein synthesis to empower the value of existing tools and expand its applications. In this sense, the combination of solid materials (normally functionalized with different biomacromolecules) together with cell-free components has made this technology more versatile and robust. In this chapter, we discuss the combination of solid materials with DNA and transcription-translation machinery to synthesize proteins within compartments, to immobilize and purify in situ the nascent protein, to transcribe and transduce DNAs immobilized on solid surfaces, and the combination of all or some of these strategies.

摘要

无细胞体系在体外生产蛋白质方面的应用彻底改变了合成生物学领域。在过去的十年中,这项技术在分子生物学、生物技术、生物医学甚至教育领域的应用不断增加。材料科学也迅速融入体外蛋白质合成领域,为现有工具赋予了新的价值,并拓展了其应用范围。从这个意义上说,将固体材料(通常用不同的生物大分子功能化)与无细胞成分相结合,使这项技术更加多功能化和稳健。在本章中,我们将讨论将固体材料与 DNA 和转录-翻译机制相结合,在隔室内合成蛋白质,就地固定和纯化新生蛋白质,在固体表面上固定和转录及转导 DNA,以及所有或部分这些策略的组合。

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