Controlling expression and inhibiting function of the toxin reporter for simple detection of the promoters’ activities in Escherichia coli was written by Chen, Yinghua;Li, Jinfeng;Zhang, Shuncheng;Hu, Jiong;Chen, Xiaofeng;Lin, Tingting;Dang, Dongya;Fan, Jun. And the article was included in Enzyme and Microbial Technology in 2022.Application In Synthesis of (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol The following contents are mentioned in the article:
The naturally occurring and mutated promoters inserted into expression plasmids or Escherichia coli chromosome are essential for recombinant protein production and metabolic engineering. Analyzing their activities and screening the promoter libraries require the simple and easy-to-use reporter. Here, we developed a novel and efficient approach to detect the promoter activity, based on E. coli cell growth inhibited by overexpression of bacteriophage ΦX174 gene E product (LyE), but recovered by pre-overexpression of Bacillus subtilis MraY (BsMraY). Under the conditional LyE construct expression in the absence or the presence of the BsMraY, activities of promoters including the reported PT7/lac, Ptac, PBAD, Prha, PhucR, PprpB, Pcum, the wild type and engineered Ptet for leaky and induced expression, the PthrC for auto-induction, and the Pms for constitutive expression were assayed. In one-plasmid coexpression system, the PBAD promoter activity detected using the reporter gene was related to the insertion site. The constructed LyE toxic effects were correlated with toxin expression levels, as determined by the split green fluorescent protein reconstitution. Microscopic anal. showed that cells lysis occurred by the LyE induced with arabinose. Taken together, the toxin reporter construct is a convenient and cost-effective tool to examine the promoter activity in E. coli. This study involved multiple reactions and reactants, such as (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1Application In Synthesis of (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol).
(2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol (cas: 367-93-1) belongs to alcohols. Under appropriate conditions, inorganic acids also react with alcohols to form esters. To form these esters, a wide variety of specialized reagents and conditions can be used. Under carefully controlled conditions, simple alcohols can undergo intermolecular dehydration to give ethers. This reaction is effective only with methanol, ethanol, and other simple primary alcohols.Application In Synthesis of (2R,3R,4S,5R,6S)-2-(Hydroxymethyl)-6-(isopropylthio)tetrahydro-2H-pyran-3,4,5-triol
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