The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.

The present disclosure provides recombinant bacterial cells that have been engineered with genetic circuitry which allow the recombinant bacterial cells to sense a patient's internal environment and respond by turning an engineered metabolic pathway on or off. When turned on, the recombinant bacterial cells complete all of the steps in a metabolic pathway to achieve a therapeutic effect in a host subject. These recombinant bacterial cells are designed to drive therapeutic effects throughout the body of a host from a point of origin of the microbiome. Specifically, the present disclosure provides recombinant bacterial cells that comprise a uric acid catabolism enzyme, e.g., a uric acid degrading enzyme, for the treatment of diseases and disorders associated with uric acid, including hyperuricemia and gout, in a subject. The disclosure further provides pharmaceutical compositions and methods of treating disorders associated with uric acid, such as hyperuricemia.

The present invention discloses a nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet and a preparation method and application thereof. Ti.sub.3C.sub.2-MXene is obtained by etching ternary layered carbides of MAX phase through hydrofluoric acid; and then, the nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet is synthesized by a simple one-step method by taking thiourea as a heteroatom source. The nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet has a unique two-dimensional layered structure, large specific surface area and abundant heteroatomic catalytic activity sites so that the material presents excellent peroxidase-like activity. The method of the present invention can successfully dope two elements of nitrogen and sulfur in one step on Ti.sub.3C.sub.2-MXene, and can effectively overcome the tedious problem of a step-by-step doping step and the secondary pollution problem of different doping sources to endow peroxidase-like activity for Ti.sub.3C.sub.2-MXene.

The present invention discloses a nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet and a preparation method and application thereof. Ti.sub.3C.sub.2-MXene is obtained by etching ternary layered carbides of MAX phase through hydrofluoric acid; and then, the nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet is synthesized by a simple one-step method by taking thiourea as a heteroatom source. The nitrogen-sulfur co-doped Ti.sub.3C.sub.2-MXene nanosheet has a unique two-dimensional layered structure, large specific surface area and abundant heteroatomic catalytic activity sites so that the material presents excellent peroxidase-like activity. The method of the present invention can successfully dope two elements of nitrogen and sulfur in one step on Ti.sub.3C.sub.2-MXene, and can effectively overcome the tedious problem of a step-by-step doping step and the secondary pollution problem of different doping sources to endow peroxidase-like activity for Ti.sub.3C.sub.2-MXene.

Embodiments of this invention disclose new second generation uric acid-sensing electrodes at least characterized by chemically bonding both uricase and the redox mediator to an electrode. The produced electrodes can be long-term stably used without losing activity. The developed electrode has been successfully applied for the analysis of uric acid (UA) in healthy human urine specimens which exhibits very good analysis accuracy and precision without too much interference. Therefore, the developed electrodes have the potential for clinical applications.

A disposable sensor chip for biological component concentration measurement includes: a chip main body defining a cavity through which a body fluid is flowable; and a reagent located in the cavity such that the body fluid flowing through the cavity comes into contact with the reagent. The reagent comprises a 2-substituted benzothiazolyl-3-substituted phenyl-5-substituted sulfonated phenyl-2H-tetrazolium salt.

A disposable sensor chip for biological component concentration measurement includes: a chip main body defining a cavity through which a body fluid is flowable; and a reagent located in the cavity such that the body fluid flowing through the cavity comes into contact with the reagent. The reagent comprises a 2-substituted benzothiazolyl-3-substituted phenyl-5-substituted sulfonated phenyl-2H-tetrazolium salt.

Methods and kits for predicting infusion reaction risk and antibody-mediated loss of response during intravenous PEGylated uricase therapy in gout patients is provided. Routine SUA monitoring can be used to identify patients receiving PEGylated uricase who may no longer benefit from treatment and who are at greater risk for infusion reactions.