Fabrication and characterization of a robust and strong bacterial promoter from a semi-rationally engineered promoter library in Bacillus subtilis |
| |
| Affiliation: | 1. School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, China;2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu, China;1. Institute of Resource Biology and Biotechnology, College of Life Science & Technology, Huazhong University of Science & Technology, Wuhan 430074, China;2. College of Life Science, South-central University for Nationalities, Wuhan 430074, China;3. Shanghai Information Center for Life Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China;4. Key Laboratory of Fermentation Engineering, Hubei University of Technology, Wuhan 430068, China;5. Department of Biotechnology and Chemical Technology, School of Chemical Technology, Aalto University, 00076 Aalto, Finland;1. Department of Integrated Biomedical and Life Sciences, Graduate School, Korea University, 145 Anam-Ro, Sungbuk-Gu, Seoul 02841, Republic of Korea;2. Department of Public Health Sciences, Graduate School, Korea University, 145 Anam-Ro, Sungbuk-Gu, Seoul 02841, Republic of Korea;3. Department of Nutritional Science and Food Management, Ewha Womans University, Seoul, 03760, Republic of Korea;4. School of Food Science and Biotechnology, Kyungpook National University, Daegu 702-701, Republic of Korea;1. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, People’s Republic of China;2. Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), No.5 Xinmofan Road, Nanjing 210009, People’s Republic of China;1. Department of Chemical Engineering, University of Waterloo, 200 University Ave West, Waterloo, ON N2L 3G1, Canada;2. Trojan Technologies, London, Ontario, Canada;1. Bioresource Utilization Laboratory, College of Engineering, China Agricultural University, Beijing, 100083 China;2. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083 China;1. Department of Food Science and Agricultural Chemistry, McGill University, 21,111 Lakeshore, Ste-Anne de Bellevue, Quebec, H9X3V9, Canada;2. Departamento de Biocatálisis. Instituto of Catálisis (CSIC), c/Marie Curie 2, 28049, Madrid, Spain |
| |
| Abstract: | Robust promoters have substantial usage in synthetic biology. The important biological components of these promoters, however, remain limited in Bacillus subtilis. In this study, an array of variant promoters derived from PsrfA was from a promoter library constructed by randomized mutation. By screening the library, the variant promoter Pv1 displayed the highest transcriptional activity, which was approximately 1.56-fold higher than that of native PsrfA. Moreover, Pv1 was able to trigger GFP expression at constantly increased levels over the culture period. The robustness of Pv1 was confirmed by the over-production of aspartase (aspA). B. subtilis that over-produced aspA exhibited normal cell growth and a constantly increased yield over the cultural period. Finally, aspA expression triggered by the Pv1 variant displayed a higher yield and expression levels of aspA in B. subtilis compared to those of native PsrfA. These results suggest that randomized mutation of sequences adjacent to the −10 region of the bacterial promoter significantly influenced the transcription activity of the promoter. Pv1, which has high activity and robustness, has potential applications in gene expression systems and genetic circuits in B. subtilis. |
| |
| Keywords: | Gene expression Promoter library Recombinant proteins Promoter engineering |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
