A Streptomyces-Based Cell-Free Protein Synthesis System for High-Level Protein Expression

doi:doi.org/10.1007/978-1-0716-1998-8_5

	A Streptomyces-Based Cell-Free Protein Synthesis System for High-Level Protein Expression
	2022-01
	Huiling, Xu; Wanqiu,Liu; Jian,Li
来源专著	Cell-Free Gene Expression: Methods and Protocols, Methods in Molecular Biology
出版地	New York
出版者	Humana New York, NY
ISBN	978-1-0716-1998-8
ISSN	1064-3745
摘要	With the rapid development of cell-free biotechnology, more and more cell-free protein synthesis (CFPS) systems have been established and optimized for protein expression in vitro. Here, we aim to improve the productivity of a newly developed Streptomyces-based CFPS system. Protein translation in CFPS systems depends on the entire endogenous translation system from cell lysates. However, lysates might lack such translation-related elements, limiting the efficiency of protein translation and therefore the productivity of CFPS systems. To address this limitation, we sought to add protein translation related factors to CFPS reactions. By doing this, the protein yield of EGFP was significantly improved up to approximately 400 μg/mL. In this chapter, we mainly describe the preparation of Streptomyces cell extracts, expression and purification of nine translation related factors, and optimization of the Streptomyces-based CFPS system for enhanced protein expression
关键词	Cell-free protein synthesis Protein expression Synthetic biology Streptomyces Translation related factors
DOI	doi.org/10.1007/978-1-0716-1998-8_5
文献类型	专著章节
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/299862
专题	物质科学与技术学院_PI研究组_李健组物质科学与技术学院_博士生
作者单位	ShanghaiTech University, School of Physical Science and Technology
第一作者单位	物质科学与技术学院
推荐引用方式 GB/T 7714	Huiling, Xu,Wanqiu,Liu,Jian,Li. A Streptomyces-Based Cell-Free Protein Synthesis System for High-Level Protein Expression. New York:Humana New York, NY,2022.