The present invention is in the field of recombinant biotechnology, in particular in the field of protein expression. The invention generally relates to methods of increasing the expression level of a protein of interest of a bacterial host cell in a production process. The invention relates particularly to improving the capacity of a bacterial host cell to express a protein of interest by expressing a phage protein during the production process which inhibits growth of the bacterial host cell. Decoupling growth of the bacterial host cell of manufacturing of the protein of interest during the production process reduces (i) the metabolic burden, (ii) oxygen demand, (iii) metabolic heat development, and (iv) avoids stress response caused by heterologous protein expression and thereby increases the capacity of a host cell to produce the protein of interest. The present invention also relates to uses of the host cell for protein expression, cell culture technology, and more specifically to culturing host cells to produce a protein of interest.

The present disclosure relates generally to bacterial delivery vehicles and their use in efficient transfer of a desired payload into a target bacterial cell of the microbiota of a subject. More specifically, the present disclosure relates to bacterial delivery vehicles with desired host ranges that can be used to efficiently transfer the desired payload in vivo to one or more target bacterial cells of the microbiota of a subject.

Some aspects of this disclosure provide methods for phage-assisted continuous evolution (PACE) of proteases. Some aspects of this invention provide methods for evaluating and selecting protease inhibitors based on the likelihood of the emergence of resistant proteases as determined by the protease PACE methods provided herein. Some aspects of this disclosure provide strategies, methods, and reagents for protease PACE, including fusion proteins for translating a desired protease activity into a selective advantage for phage particles encoding a protease exhibiting such an activity and improved mutagenesis-promoting expression constructs. Evolved proteases that recognize target cleavage sites which differ from their canonical cleavage site are also provided herein.

The present disclosure is directed to a universal antivenom for the treatment of venomous animal bites, and methods of developing the same using a novel targeted phage display technique.

The present disclosure is directed to a universal antivenom for the treatment of venomous animal bites, and methods of developing the same using a novel targeted phage display technique.

The disclosure relates to genetic engineered bacteria having a genetic memory circuit, compositions thereof, formulations thereof, methods of analyses and method of treatment of conditions related to the gastrointestinal tract including the mouth and the stomach.

The disclosure relates to genetic engineered bacteria having a genetic memory circuit, compositions thereof, formulations thereof, methods of analyses and method of treatment of conditions related to the gastrointestinal tract including the mouth and the stomach.

A protein expression system for use in a prokaryotic host is provided, the expression system comprising: a) an expression cassette comprising a nucleic acid sequence encoding a protein of interest operably linked to a T7 RNA polymerase-dependent promoter; and b) an expression cassette comprising a nucleic acid sequence encoding T7 RNA polymerase operably linked to a host polymerase-dependent k phage promoter and a single perfect palindrome operator sequence; wherein the expression cassette for T7 RNA polymerase is located on the chromosome of a host cell.

A protein expression system for use in a prokaryotic host is provided, the expression system comprising: a) an expression cassette comprising a nucleic acid sequence encoding a protein of interest operably linked to a T7 RNA polymerase-dependent promoter; and b) an expression cassette comprising a nucleic acid sequence encoding T7 RNA polymerase operably linked to a host polymerase-dependent k phage promoter and a single perfect palindrome operator sequence; wherein the expression cassette for T7 RNA polymerase is located on the chromosome of a host cell.

The present invention relates to a vector, preferably included in a delivery vehicle, comprising no more than 100, preferably no more than 10, restriction sites recognized by the restriction enzymes encoded by each bacterium of a group of bacteria of interest. The invention also relates to the use of said vector, preferably included in a delivery vehicle, as a drug, especially in the treatment of a disease in a patient in need thereof.