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 to enzyme inhibitors. More specifically, the present invention relates to ligand-directed covalent modification of proteins; method of designing same; pharmaceutical formulation of same; and method of use.

Provided is a method for producing 1,3-propanediol by means of fermentation of a recombinant microorganism. First, a recombinant microorganism is provided; the recombinant microorganism can overexpress acetyl-CoA carboxylase genes: accBC and accDA, a malonyl-CoA synthetase gene: mcr, a 3-hydroxypropionyl-CoA synthetase gene: pcs, a 3-hydroxypropionyl-CoA reductase gene: pduP, and a 1,3-propanediol reductase gene: yqhD. The recombinant microorganism is subjected to fermentation culture in a flask or fermentor using glucose ad as raw material to obtain the 1,3-propanediol. The recombinant microorganism can utilize low-cost glucose, sucrose, molasses, xylose and the like as raw material in the fermentation process, without additional expensive vitamin B12. Thus, cost of the production is significantly reduced, and there is a promising prospect in market.

Disclosed are gene engineering bacteria for producing L-arginine and a construction method and an application of the gene engineering bacteria. According to the method, genes encoding a carbamoyl phosphate synthetase and a gene encoding an L-arginine biosynthesis pathway enzyme are integrated into Escherichia coli; the present invention has analyzed and reconstructed the arginine synthetic pathway and the metabolic flow related to arginine in the entire amino acid metabolic network in E. coli and finally obtained a genetically engineered bacterial strain which has a clear genetic background, carries no plasmids, undergoes no mutagenesis and is capable of stably and efficiently producing L-arginine.

The present disclosure relates to methods of synthesizing moderate length RNAs by splint-mediated ligation of RNA fragments.

Compositions and methods for detecting nucleic acid-protein interactions, or more generally interactions between a nucleic acid and another molecule. A Cas protein (e.g., a catalytically dead Cas13) is fused to a proximity tagging enzyme (e.g., a Pup ligase) and thus brings the proximity tagging enzyme to the proximity of a protein that binds to a nucleic acid, when the Cas protein recognizes the nucleic acid, e.g., through a guide RNA. The proximity tagging enzyme then tags the protein enabling it to be identified as a protein that interacts with the nucleic acid.

Angelman Syndrome (AS) is a genetic disorder occurring in approximately one in every 15,000 births. It is characterized by severe mental retardation, seizures, difficulty speaking and ataxia. The gene responsible for AS was discovered to be UBE3A and encodes for E6-AP, an ubiquitin ligase. A unique feature of this gene is that it undergoes maternal imprinting in a neuron-specific manner. In the majority of AS cases, there is a mutation or deletion in the maternally inherited UBE3A gene, although other cases are the result of uniparental disomy or mismethylation of the maternal gene. While most human disorders characterized by severe mental retardation involve abnormalities in brain structure, no gross anatomical changes are associated with AS. We have generated a Ube3a protein with additional sequences that should allow the secretion from cells and uptake by neighboring neuronal cells. This would confer a functional E6-AP protein into the neurons and rescue disease pathology.

The present invention relates to the use of glutamine synthetase as a protein therapy (such as enzyme replacement protein therapy) for the treatment of hyperammonemia. In particular the invention relates to the systemic administration of glutamine synthetase. The glutamine synthetase may be provided in conjugated or fusion form, to increase its half-life in the circulation. Also provided is a pharmaceutical composition comprising glutamine synthetase. The invention also relates to the uses, methods, and compositions involving a combination of the glutamine synthetase protein and an ammonia lowering agent, such as a nitrogen scavenger.

The invention relates to a system for expressing nonribosomal peptide synthetases (NRPSs), polyketide synthases (PKS) or NRPS/PKS hybrid synth(et)ases. NRPS, PKS or hybrids thereof are large multi-domain proteins or multi-domain complexes, the expression of which for the production of peptides often causes difficulties. The invention correspondingly relates to a system for expressing portions of the enzymes which can be assembled post-translationally via protein-protein interactions, introduced in a targeted manner, to form multi-enzyme complexes. The invention discloses protein fragments of such an assembly, and the nucleic acids coding therefor. The invention also relates to a vector system for the protein fragments of the invention and its use for producing functional NRPS/PKS enzyme complexes.

The invention relates generally to methods of producing a protein comprising a first unnatural amino acid (UAA) and a second, different UAA, and proteins comprising a first UAA and a second, different UAA.