Professor Liu Long's team from Jiangnan University Recently "Nucleic Acids Research" (IF16.6) The article "A cross-species inducible system for enhanced protein expression and multiplexed metabolic pathway fine-tuning in bacteria" was published in the journal. Qihengxing's flagship product StarLighter HotStart Taq Pro PCR Mix (FS-P5001, StarLighter Hot Start Taq Pro PCR Master Mix) I am fortunate to be able to participate in this and contribute to this research!
In this study, researchers selected nine reported induction systems, including IPTG and xylose. Through rational design and random mutagenesis, they reconfigured these systems to achieve efficient expression in diverse strains. By introducing a mutant suppressor expression library, they further optimized the induction systems to reduce leaky expression. The induction systems were then placed on plasmids and genomes, and their expression performance was tested in Escherichia coli, Bacillus subtilis, and Corynebacterium glutamicum. Ultimately, two cross-species induction systems were successfully constructed: the 2,4-diacetylphloroglucinol (DAPG)-inducible system PphlF3R1 and the anhydrotetracycline (aTc)-inducible system Ptet2R2*. After optimization, Ptet2R2* demonstrated low leakiness, a wide dynamic range, sufficient expression intensity, and appropriate sensitivity in all three strains. This system was used to effectively regulate the expression of various reporter proteins (sfGFP, mCherry, and mScarlet3) and gene clusters (crtEIB, crtEIBY, and vioABCDE). Furthermore, a single-input gene circuit based on T7 RNA polymerase (T7 RNAP) and dCas12a was developed to simultaneously activate and repress gene expression.
