One-step cloning and targeted duplication of Pantoea ananatis chromosomal fragments.

In this study, we describe a novel method for one-step cloning and targeted duplication of P. ananatis chromosomal fragments. According to this method, the chromosomal region of interest is subcloned in vivo via λ Red recombination into the short synthetic non-replicable DNA fragment containing the excisable antibiotic-resistance marker gene and φ80 att-P site.

H-Translocating Membrane-Bound Pyrophosphatase from Fuels Cells via an Alternative Pathway for Energy Generation.

Inorganic pyrophosphatases (PPases) catalyze an essential reaction, namely, the hydrolysis of PP, which is formed in large quantities as a side product of numerous cellular reactions. In the majority of living species, PP hydrolysis is carried out by soluble cytoplasmic PPase (S-PPases) with the released energy dissipated in the form of heat. In part of this energy can be conserved by proton-pumping pyrophosphatase (H-PPase) in the form of a proton electrochemical gradient for further ATP synthesis.

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.

The Amino Acid Uptake Protein CycA: Regulation of Its Synthesis and Practical Application in l-Isoleucine Production.

Amino acid transport systems perform important physiological functions; their role should certainly be considered in microbial production of amino acids. Typically, in the context of metabolic engineering, efforts are focused on the search for and application of specific amino acid efflux pumps. However, in addition, importers can also be used to improve the industrial process as a whole.

A novel one-step method for targeted multiplication of DNA fragments from the Escherichia coli chromosome mediated by coordinated functioning of λ and φ80 bacteriophage recombination systems.

A novel technique for targeted stable multiplication of a specific long E. coli chromosome fragment was developed. The method is based on the coordinated functioning of λ and φ80 bacteriophage site-specific recombination and integration systems.