The present invention provides an application of a cold-active bacteriophage protein in a room temperature nucleic acid amplification reaction; the cold-active bacteriophage is selected from vB_EcoM-VR5, vB_EcoM-VR7, and vB_EcoM-VR20,vB_EcoM-VR25, or vB_EcoM-VR26, and the cold-active bacteriophage protein is a uvsX protein, a uvsY protein and a gp32 protein and/or a variant protein having corresponding functions. Preferably, the uvsX protein and the variant protein thereof are selected from any sequence of SEQ ID Nos. 1-23 or 30, the uvsY protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.27-29 or 32, and the gp32 protein and the variant protein thereof are selected from any sequence of SEQ ID Nos.24-26 or 31. The present invention further provides a room temperature nucleic acid amplification reaction system containing the cold-active bacteriophage protein.

Methods of expressing an expression product of interest are provided. Accordingly there is provided a method comprising introducing into a cell a polynucleotide comprising an AimR responsive element operatively linked to a nucleic acid sequence encoding the expression product of interest, and contacting said cell with an AimP peptide comprising an amino acid sequence of XXXXGG/A, wherein said AimP peptide is capable of binding said AimR polypeptide and dissociating said AimR polypeptide from said AimR responsive element. Also provided are articles of manufacture, isolated peptides, polynucleotides and nucleic acid constructs.

Provided are a novel polypeptide having endolysin activity, a fusion protein comprising the polypeptide and an antibiotic active protein, and an antibiotic use against a gram-negative pathogen of the polypeptide and/or fusion protein and/or a use for prevention and/or treatment of gram-negative pathogen infection and/or disease or symptoms related to gram negative pathogen infection.

Disclosed here are phage compositions comprising CRISPR-Cas systems and methods of use thereof. In certain embodiments, disclosed herein is a bacteriophage comprising a nucleic acid sequence encoding a Type I CRISPR-Cas system comprising: (a) a CRISPR array; (b) a Cascade polypeptide; and (c) a Cas3 polypeptide. In some embodiments, the CRISPR array comprises a spacer sequence and at least one repeat sequence. In some embodiments, the at least one repeat sequence is operably linked to the spacer sequence at either its 5′ end or its 3′ end.

The present disclosure provides a chimeric receptor binding protein (RBP) resistant to proteolytic digestion wherein said RBP comprises a portion of a receptor binding protein derived from a bacteriophage fused through a designed linker region consisting of 1 to 70 amino acids, to a portion of a receptor binding protein derived from a different bacteriophage, wherein said linker region is designed to be resistant to proteolytic digestion.

The present invention relates to an antibacterial protein EFAL-2 derived from bacteriophage Ent-FAP-4 (Accession number: KCTC 12854BP), which has the ability to kill Enterococcus faecium and an amino acid sequence represented by SEQ ID NO: 2, a pharmaceutical composition containing the same as an active ingredient, and a method for preventing or treating diseases caused by Enterococcus faecium using the pharmaceutical composition.

There is disclosed a method of engineering bacteriophages comprising: identifying a bacteriophage with only one attachment gene; isolating said bacteriophage; removing said attachment gene from the genome of said bacteriophage; and inserting a non-natural attachment gene into the genome of said bacteriophage wherein said non-natural attachment gene is specific for attaching to a selected bacteria. There is also disclosed a mutant bacteriophage comprising a heterologous nucleic acid sequence encoding a first specific attachment gene, the first specific attachment gene being different than an inactivated attachment gene and being specific for a selected bacteria. In another embodiment, there is disclosed a method of eliminating a microbial contaminant, the method comprising: obtaining one or more lytic enzymes produced by a mutant bacteriophage; applying the one or more lytic enzymes to a bacterial contaminant, without prior infection of the bacterial contaminant with a bacteriophage, to eliminate the bacterial contaminant.

The present invention relates to new species-selective phage endolysins and their use to treat bacterial vaginosis (BV). The present invention provides recombinant endolysins, i.e. domain-swapped endolysins. The invention also relates to said endolysins for use in treating diseases or disorders such as bacterial infections, in particular BV. The invention further relates to polynucleotides encoding said endolysins. Said polynucleotides can also be used for treating such diseases or disorders. Also provided by the present invention is a pharmaceutical composition comprising an endolysin of the invention for use in treating such diseases or disorders. Said endolysins, polynucleotides and pharmaceutical composition may be administered locally, in particular locally into the vagina.

Hybrid nanopores, comprising a protein pore supported within a solid-state membrane, which combine the robust nature of solid-state membranes with the easily tunable and precise engineering of protein nanopores. In an embodiment, a lipid-free hybrid nanopore comprises a water soluble and stable, modified portal protein of the Thermus thermophilus bacteriophage G20c, electrokinetically inserted into a larger nanopore in a solid-state membrane. The hybrid pore is stable and easy to fabricate, and exhibits low peripheral leakage, allowing sensing and discrimination among different types of biomolecules.

The present invention is in the field of recombinant biotechnology, in particular in the field of protein expression and nucleotide production. The invention generally relates to systems and processes that are suitable or comprise a two stage production process, in which the growth of the bacterial host cell is spatiotemporally separated from the production of the protein or nucleic acid of interest. Accordingly, the present invention relates to a system and a process for use in continuous production of a protein of interest or a nucleotide of interest by a bacterial host cell.