Systematic identification of a panel of strong promoter regions from Listeria monocytogenes for fine-tuning gene expression

Microbial Cell Factories

Junfei Ma¹  Qianyu Ji¹  Shuying Wang¹  Qing Liu¹ 

[1] School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, 200093, Shanghai, People’s Republic of China;

关键词: Listeria monocytogenes;    Promoters;    RNA-seq;    Exogenous protein;   

DOI  :  10.1186/s12934-021-01628-w

来源: Springer

【 摘 要 】

BackgroundAttenuated Listeria monocytogenes (Lm) has been widely used as a vaccine vector in the prevention and treatment of pathogen infection and tumor diseases. In addition, previous studies have proved that the attenuated Lm can protect zebrafish from Vibrio infections, indicating that the attenuated Lm has a good application prospect in the field of aquatic vaccines. However, the limitation mainly lies in the lack of a set of well-characterized natural promoters for the expression of target antigens in attenuated Lm.ResultsIn our study, candidate strong promoters were identified through RNA-seq analysis, and characterized in Lm through enhanced green fluorescent protein (EGFP). Nine native promoters that showed stronger activities than that of the known strong promoter P36 under two tested temperatures (28 and 37 °C) were selected from the set, and P29 with the highest activity was 24-fold greater than P36. Furthermore, we demonstrated that P29 could initiate EGFP expression in ZF4 cells and zebrafish embryos.ConclusionsThis well-characterized promoter library can be used to fine-tune the expression of different proteins in Lm. The availability of a well-characterized promoter toolbox of Lm is essential for the analysis of yield increase for biotechnology applications.

【 授权许可】

CC BY