Abstract
The ease of genetic manipulation, low cost, rapid growth and number of previous studies have made Escherichia coli one of the most widely used microorganism species for producing recombinant proteins. In this post-genomic era, challenges remain to rapidly express and purify large numbers of proteins for academic and commercial purposes in a high-throughput manner. In this review, we describe several state-of-the-art approaches that are suitable for the cloning, expression and purification, conducted in parallel, of numerous molecules, and we discuss recent progress related to soluble protein expression, mRNA folding, fusion tags, post-translational modification and production of membrane proteins. Moreover, we address the ongoing efforts to overcome various challenges faced in protein expression in E. coli, which could lead to an improvement of the current system from trial and error to a predictable and rational design.
Highlights
High-throughput studies can be defined as research that allows thousands of concurrent measurements of biological molecules to be obtained and makes large-scale repetition feasible
Successful recombinant protein expression and purification is frequently indispensable for both basic research studies and biotechnological and commercial applications [166]
Highthroughput protein expression and purification in E. coli has begun to revolutionize the manner in which studies are conducted in various research fields
Summary
High-throughput studies can be defined as research that allows thousands of concurrent measurements of biological molecules to be obtained and makes large-scale repetition feasible. This technology originated in the early 1990s when the first automated DNA sequencers were developed and human genome sequencing was initiated [1]. A parallel and high-throughput approach must be employed in protein expression and purification, which has been the bottleneck in studies of protein function, structure and application in the post-genomic era [6]. Numerous advances in these methods have been made over the past few years, in this review, we discuss the advantages and disadvantages of the current methods—those targeting gene cloning, vector construction, fusion tags and host strains
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
