A new phagemid cloning vector for positive selection of recombinants, pBa-7, was constructed which contains an active barnase gene encoding the cytotoxic ribonuclease from Bacillus amyloliquefaciens, under control of the lac promoter. PBa-7 is a derivative of the high-copy number pBluescript II KS+ phagemid in which the modified barnase killer gene has been fused downstream from the lac promote...

The amino acid L-serine is required for pharmaceutical purposes, and the availability of a sugar-based microbial process for its production is desirable. However, a number of intracellular utilization routes prevent overproduction of L-serine, with the essential serine hydroxymethyltransferase (SHMT) (glyA) probably occupying a key position. We found that constructs of Corynebacterium glutamicu...

Crude bacterial cell extracts prepared from an Escherichia coli lacIq strain were shown to protect specific nucleotides in the lac operator from methylation by dimethyl sulfate (DMS) or digestion by DNase I, whereas no protection was observed using extracts prepared from a nearly isogenic lacI- strain. These experiments show that it is not necessary to use purified regulatory proteins in experi...

We have constructed two expression vectors based on the pJF118HE vector developed for Escherichia coli by Fürste et al. [Gene 48 (1986) 119-131]. The tac promoter (ptac) was exchanged for the trc promoter (ptrc) and an NdeI site was created at the appropriate distance from the ribosome-binding site. The NdeI site permits cloning of a gene at its translation start point without altering the amin...

This work demonstrates improvement of a whole-cell cadmium detection sensor through construction of a gene circuit. A cadmium (II) specific regulatory promoter, P(cadR,) from Psuedomonas putida 06909, is used in the assembly of a toggle circuit. The circuit contains the cadR promoter fused to lacIq and gfp, and a divergently transcribed tac promoter and cadR. The toggle sensor exhibits lower ba...

Crosstalk between fluorescent proteins spectra might be an issue regarding systems containing both GFP-and RFP-expressing devices. In order to validate whether this phenomenon was significantly present with the acquisition system used in this study (i.e., the Infinite F200 microplate reader, Tecan), at least in the tested conditions for the combined gene expression cassettes, an ad-hoc experime...

Trehalose (α-D-glucopyranosyl-1,1-α-D-glucopyranoside) is a stable and nonreducing disaccharide; can be used as sweetener, stabilizer, humectant; has many applications in the food, pharmaceutical, cosmetic industries. production from maltose catalyzed by trehalose synthase (TreS) simple economically feasible for industrial-scale application. Reducing cost enhancing efficiency of TreS synthesis ...

In vivo and in vitro recombination methods were used to construct the recombinant plasmid pTG801, in which the F-plasmid DNA transfer (tra) genes required for the formation of functional F pili were placed under the lac transcriptional control sequences of pUC19. The 20 kilobases of cloned F DNA includes genes traA through the 5'-terminal part of traG; the plasmid lacks the positive regulatory ...

A novel cross-regulation system employing two pairs of interacting promoter-repressor systems was constructed using the tac-lacI and lambda pL-cI promoter-operator-repressor systems. In particular, transcription of the cat gene and the fused cI gene is regulated by the tac promoter, while transcription of the lacI gene is controlled by the lambda pL promoter. In order to compare CAT production ...