Selection of endogenous expression elements from Corynebactrium glutamicum

Abstract:

[Objective] We constructed a Corynebacterium glutamicum expression-element probe vector and selected sequences that can activate expression of protein. [Methods] Based on the classic plasmid pXMJ19, we set the inserted position with the new method of Golden Gate cloning strategy that can connect material sequence to the report gene seamlessly, avoiding remaining unwanted sequence that may interfere the measurement of expression elements effect. By analyzing the transcriptomic data of C. glutamicum BZH001 cultivated under different dissolved oxygen level in our previous study, we screened six genes stabilizing in high transcriptional level, analyzed their promoter regions and 5′UTR regions by the promoter software prediction, and then amplified the promoter sequences associated with 5′UTR regions of the six genes. The activities of these expression elements were measured using the reporter gene egfp in the probe vector. [Results] Five expression elements with different performances were achieved, and the best one could reach over 3 500 RFU/OD600 of fluorescent intensity. [Conclusion] Associated with the transcriptomic data, we can screen effective expression elements using this probe vector, which can provide more genetic parts for the future genetic engineering and construction of biological system in Corynebacterium glutamicum.

Key words:Corynebacterium glutamicum, RNA-Seq, Promoter, Probe-vector