Construction of a High-Expression System in Bacillus through Transcriptomic Profiling and Promoter Engineering.

Microorganisms

Key Laboratory of Molecular Biology and Biotechnology, College of Life Sciences, Sichuan University, 29, Wangjiang Rd, Chengdu 610064, China.

Published: July 2020


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Article Abstract

is an ideal host for secretion and expression of foreign proteins. The promoter is one of the most important elements to facilitate the high-level production of recombinant protein. To expand the repertoire of strong promoters for biotechnological applications in species, 14 highly transcribed genes based on transcriptome profiling of BA06 were selected and evaluated for their promoter strength in . Consequently, a strong promoter P was obtained, which could drive the genes encoding alkaline protease () and green fluorescent protein (GFP) to express more efficiency by an increase of 3.65-fold and 18.40-fold in comparison with the control promoter (P), respectively. Further, promoter engineering was applied to P, leading to a mutation promoter (P) that could increase GFP expression by 3.67-fold over the wild-type promoter (P). Moreover, the IPTG-inducible expression systems were constructed using the operon based on the strong promoters of P and P, which could work well both in and . In this study, highly efficient expression system for was constructed based on transcriptome data and promoter engineering, which provide not only a new option for recombinant expression in , but also novel genetic tool for .

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7409208PMC
http://dx.doi.org/10.3390/microorganisms8071030DOI Listing

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