A highly efficient targeted recombination system for engineering linear chromosomes of industrial bacteria Streptomyces.

Soil bacteria Streptomyces are the most important producers of secondary metabolites, including most known antibiotics. These bacteria and their close relatives are unique in possessing linear chromosomes, which typically harbor 20 to 30 biosynthetic gene clusters of tens to hundreds of kb in length. Many Streptomyces chromosomes are accompanied by linear plasmids with sizes ranging from several to several hundred kb.

Is Required for Intracellular Replication of in Various Human Cells and Affects Its Polyamine Metabolism and Global Transcriptomes.

The gene has been reported to regulate polyamine metabolism in and , but its role in remains unknown. Our preliminary studies have revealed that widely affects the transcriptomes of infected M and Caco-2 cells and that it is required for the intracellular replication of serovar Typhimurium (. Typhimurium) in HeLa cells.

Telomere associated primase Tap repairs truncated telomeres of Streptomyces.

Replication of the linear chromosomes of soil bacteria Streptomyces proceeds from an internal origin towards the telomeres, followed by patching of the resulting terminal single-strand overhangs by DNA synthesis using terminal proteins as the primer, which remains covalently bound to the 5΄ ends of the DNA. In most Streptomyces chromosomes, the end patching requires the single-strand overhangs, terminal protein Tpg, and terminal associated protein Tap. The telomere overhangs contain several palindromic sequences capable of forming stable hairpins.

Micro-RNA quantification using DNA polymerase and pyrophosphate quantification.

A rapid quantification method for micro-RNA based on DNA polymerase activity and pyrophosphate quantification has been developed. The tested micro-RNA serves as the primer, unlike the DNA primer in all DNA sequencing methods, and the DNA probe serves as the template for DNA replication. After the DNA synthesis, the pyrophosphate detection and quantification indicate the existence and quantity of the tested miRNA.