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Construction of a High-Expression System in Bacillus through Transcriptomic Profiling and Promoter Engineering
Cui-Cui Miao1, Lin-Li Han1, Yan-Bing Lu1
1Key Laboratory of Molecular Biology and Biotechnology, College of Life Sciences, Sichuan University, 29, Wangjiang Rd, Chengdu 610064, China.
Microorganisms
|July 16, 2020
Summary
Researchers identified a strong promoter, P2069, from Bacillus pumilus for enhanced recombinant protein production in Bacillus subtilis. Promoter engineering further improved expression, creating a valuable tool for biotechnology.
Area of Science:
- Microbial biotechnology
- Molecular biology
- Genetics
Background:
- Bacillus subtilis is a key host for foreign protein expression.
- Strong promoters are crucial for high-level recombinant protein production.
- Expanding the library of robust Bacillus promoters is essential for biotechnological advancements.
Purpose of the Study:
- To identify and characterize novel strong promoters from B. pumilus for use in B. subtilis.
- To engineer existing strong promoters for even higher expression levels.
- To develop inducible expression systems for B. subtilis and B. pumilus.
Main Methods:
- Transcriptome profiling of B. pumilus BA06 to identify highly transcribed genes.
- Evaluation of promoter strength in B. subtilis using alkaline protease (aprE) and green fluorescent protein (GFP) reporter genes.
- Promoter engineering to enhance promoter activity.
- Construction of IPTG-inducible expression systems.
Main Results:
- A novel strong promoter, P2069, was identified, increasing aprE and GFP expression by 3.65-fold and 18.40-fold, respectively, compared to the control.
- Promoter engineering yielded P2069M, further boosting GFP expression by 3.67-fold.
- IPTG-inducible expression systems based on P2069 and P2069M demonstrated efficacy in both B. subtilis and B. pumilus.
Conclusions:
- A highly efficient expression system for Bacillus was developed using transcriptome data and promoter engineering.
- The novel promoters P2069 and P2069M offer improved recombinant protein expression in B. subtilis.
- These findings provide a new genetic tool for B. pumilus and enhance options for B. subtilis recombinant expression.
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