Zr-MOFs was applied for the immobilization of hyperthermophilic and halophilic amino acid dehydrogenase (Zr-MOFs-NTAaDH) by physical adsorption for the biosynthesis of L-homophenylalanine. Activity of Zr-MOFs-NTAaDH was enhanced by 3.3-fold of the free enzyme at 70°C.
View Article and Find Full Text PDFSheng Wu Gong Cheng Xue Bao
April 2023
Manipulation of genes, including knock-out or knock-in, replacement of gene elements (such as promoters), fusion with a fluorescent protein gene, and construction of gene reporter, is required in most of the biotechnological laboratories. The widely used gene manipulating methods based on two-step allelic exchange are cumbersome in terms of constructing plasmids, transforming and screening. In addition, the efficiency of using this method for long fragment knockout is low.
View Article and Find Full Text PDFMicrobiol Spectr
February 2023
Transcriptional factors (TFs) and their regulons make up the gene regulatory networks. Here, we developed a method based on TF-directed activation-induced cytidine deaminase (AID) mutagenesis in combination with genome sequencing, called AIDmut-Seq, to detect TF targets on the genome. AIDmut-Seq involves only three simple steps, including the expression of the AID-TF fusion protein, whole-genome sequencing, and single nucleotide polymorphism (SNP) profiling, making it easy for junior and interdisciplinary researchers to use.
View Article and Find Full Text PDFThe part of expression noise that is brought about by transcriptional regulation (represented here as NTR) is an important criterion for estimating the regulatory mode of a gene. However, characterization of NTR is an under-explored area, and there is little knowledge regarding the genome-wide NTR in the model pathogen Pseudomonas aeruginosa. Here, with a library of dual-color transcriptional reporters, we estimated the NTR for over 90% of the promoters in P.
View Article and Find Full Text PDFQuantification of the rate of direct repeat deletion (DRD) is an important aspect in the research of DNA rearrangement. The widely used tetracycline selection method usually introduces antibiotic pressure to the tested organism, which may interfere with the DRD process. Also the length of repeat arm (LRA) is limited by the length of the TetR coding sequence.
View Article and Find Full Text PDFEnvironmental bacteria persistently exist in hospitals and thereby often contaminate biomedical devices, which usually causes device-associated infections that have become a major cause of patient illness and death in the hospital. In this study, for the first time, the identification of strong shear flow persister (SSP) cells in Pseudomonas aeruginosa is reported. Unlike common persister cells that are highly tolerant to antibiotics, it is reported that the SSP cells can resist mechanical washings on the surfaces of various polymer materials and can form distinctive biofilms that are tolerant to high doses of aminoglycoside antibiotics.
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