Team of Professor Liu Long 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 Qihengxing's flagship product StarLighter HotStart Taq Pro PCR Mix (FS-P5001, StarLighter Hot Start Taq Pro PCR premix) I am fortunate to be able to participate in this and contribute to this research!
In the article, the researchers selected 9 reported induction systems, including IPTG induction system and xylose induction system. These systems were reconstructed through rational design and random mutation to enable effective expression in different strains. The induction system was further optimized by introducing a mutant suppressor expression library to reduce leaky expression. The induction system was placed on plasmids and genomes to test its expression performance in Escherichia coli, Bacillus subtilis and Corynebacterium glutamicum. Finally, two cross-species induction systems were successfully constructed: 2,4-diacetylphloroglucinol (DAPG) induction system PphlF3R1 and anhydrotetracycline (aTc) induction system Ptet2R2*. Through optimization, P_tet2R2* showed low leakage, 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). In addition, a single-input genetic circuit based on T7 RNA polymerase (T7 RNAP) and dCas12a was developed for simultaneous activation and repression of gene expression.
