Genome engineering of the Corynebacterium glutamicum chromosome by the Extended Dual-In/Out strategy.

A novel genome editing method for repeated introduction of foreign DNA, including insertion of rather large DNA fragments, into predesigned points in the Corynebacterium glutamicum chromosome was developed. The method is based on the implementation of the Dual-In/Out strategy, which was previously provided in Escherichia coli according to recombineering-based methods (Minaeva et al., 2008) and allowed step-by-step construction of marker-less plasmid free recombinant strains.

Efficient production of long double-stranded RNAs applicable to agricultural pest control by Corynebacterium glutamicum equipped with coliphage T7-expression system.

RNA-based pesticides exert their function by suppressing the expression of an essential gene in the target pest through RNA interference caused by double-stranded RNA (dsRNA). Here, we selected target genes for growth suppression of the solanaceous crop pests ladybird beetle (Henosepilachna vigintioctopunctata) and Colorado potato beetle (Leptinotarsa decemlineata)-the death-associated inhibitor of apoptosis protein 1 gene (diap1), and an orthologous gene of the COPI coatomer protein complex (copI), respectively. We constructed a cost-competitive overproduction system for dsRNA using Corynebacterium glutamicum as a host bacterium.

Universal Actuator for Efficient Silencing of Genes Based on Convergent Transcription Resistant to Rho-Dependent Termination.

Dynamic control is a distinguished strategy in modern metabolic engineering, in which inducible convergent transcription is an attractive approach for conditional gene silencing. Instead of a simple strong "reverse" () promoter, a three-component actuator has been developed for constitutive genes silencing. These actuators, consisting of promoters with different strengths, the ribosomal transcription antitermination-inducing sequence -AT, and the RNase III processing site, were inserted into the 3'-UTR of three metabolic genes.

Specific features of L-histidine production by Escherichia coli concerned with feedback control of AICAR formation and inorganic phosphate/metal transport.

Background: In the L-histidine (His) biosynthetic pathway of Escherichia coli, the first key enzyme, ATP-phosphoribosyltransferase (ATP-PRT, HisG), is subject to different types of inhibition. Eliminating the feedback inhibition of HisG by the His end product is an important step that enables the oversynthesis of His in breeding strains. However, the previously reported feedback inhibition-resistant mutant enzyme from E.

Mu-driven transposition of recombinant mini-Mu unit DNA in the Corynebacterium glutamicum chromosome.

A dual-component Mu-transposition system was modified for the integration/amplification of genes in Corynebacterium. The system consists of two types of plasmids: (i) a non-replicative integrative plasmid that contains the transposing mini-Mu(LR) unit bracketed by the L/R Mu ends or the mini-Mu(LER) unit, which additionally contains the enhancer element, E, and (ii) an integration helper plasmid that expresses the transposition factor genes for MuA and MuB. Efficient transposition in the C.

Gyroscopic effect detection in the colliding-pulse hybridly mode-locked erbium-doped all-fiber ring soliton laser.

We report on the gyroscopic effect detection in the bidirectional ultra-short pulse hybridly mode-locked erbium-doped all-fiber ring soliton laser. Owing to the Kerr nonlinearity contribution through self-phase modulation and self-steepening effects to the carrier-to-envelope phase slip of both clockwise and counterclockwise solitons, wideband controllable tuning of a gyroscope bias point has been demonstrated by means of appropriate adjustment of either intracavity polarization or pump power. Angular velocity detected ranges from 0.

Complete nucleotide sequence and annotation of the temperate corynephage ϕ16 genome.

The complete genome of ϕ16, a temperate corynephage from Corynebacterium glutamicum ATCC 21792, was sequenced and annotated (GenBank: KY250482). The electron microscopy study of ϕ16 virion confirmed that it belongs to the family Siphoviridae. The ϕ16 genome consists of a linear double-stranded DNA molecule of 58,200 bp (G+C = 52.

Generation regimes of bidirectional hybridly mode-locked ultrashort pulse erbium-doped all-fiber ring laser with a distributed polarizer.

We report on the stable picosecond and femtosecond pulse generation from the bidirectional erbium-doped all-fiber ring laser hybridly mode-locked with a coaction of a single-walled carbon nanotube-based saturable absorber and nonlinear polarization evolution that was introduced through the insertion of the short-segment polarizing fiber. Depending on the total intracavity dispersion value, the laser emits conservative solitons, transform-limited Gaussian pulses, or highly chirped stretched pulses with almost 20 nm wide parabolic spectrum in both clockwise (CW) and counterclockwise (CCW) directions of the ring. Owing to the polarizing action in the cavity, we have demonstrated for the first time, to the best of our knowledge, an efficient tuning of soliton pulse characteristics for both CW and CCW channels via an appropriate polarization control.

High-energy, sub-100 fs, all-fiber stretched-pulse mode-locked Er-doped ring laser with a highly-nonlinear resonator.

We report on ultra-short stretched pulse generation in an all-fiber erbium-doped ring laser with a highly-nonlinear germanosilicate fiber inside the resonator with a slightly positive net-cavity group velocity dispersion (GVD). Stable 84 fs pulses were obtained with a 12 MHz repetition rate at a central wavelength of 1560 nm with a 48.1 nm spectral pulse width (full width at half maximum, FWHM) and 30 mW average output power; this corresponds to the 29.

Oligonucleotide recombination in corynebacteria without the expression of exogenous recombinases.

Brevibacterium lactofermentum and Corynebacterium glutamicum are important biotechnology species of the genus Corynebacterium. The single-strand DNA annealing protein (SSAP)-independent oligonucleotide-mediated recombination procedure was successfully applied to the commonly used wild-type strains B. lactofermentum AJ1511 and C.

Pho regulon promoter-mediated transcription of the key pathway gene aroGFbr improves the performance of an L-phenylalanine-producing Escherichia coli strain.

DAHP synthase (EC 4.1.2.

Conditional silencing of the Escherichia coli pykF gene results from artificial convergent transcription protected from Rho-dependent termination.

PykF is one of two pyruvate kinases in Escherichia coli K-12. lambdaP(L) was convergently integrated into the chromosome of the MG1655 strain, downstream of pykF, face-to-face with its native promoter. In the presence of lambdacIts857, efficient pykF ts-silencing was achieved when the 5'-terminus of the P(L)-originated antisense RNA (asRNA), consisting of the rrnG-AT sequence, converted elongation complexes of RNA polymerase to a form resistant to Rho-dependent transcription termination.