Abstract: The present invention relates to immunogenic compositions, such as vaccines, comprising immunogenic polypeptides from Haemophilus influenzae and Moraxella catarrhalis, for use in methods of boosting an immune response and methods of treatment using same. More particularly, the invention relates to use of such immunogenic compositions in methods of treating or preventing exacerbation of chronic obstructive pulmonary disease.

Abstract: Provided are a composition for a polymerase reaction, containing a nucleic acid polymerase and a 2-methacryloyloxyethyl phosphorylcholine (MPC)-containing zwitterionic copolymer detergent, a tube for a polymerase reaction, and a kit for a polymerase reaction. The stability of the composition for a polymerase reaction can be improved and the reliability of the results of polymerase reaction such as nucleic acid polymerization or amplification can be improved.

Abstract: The present invention relates to a method of producing a recombinant protein in a host cell comprising adding Polyethyleneimine (PEI) during cell culture. Addition of PEI to the cell culture as a fermentation enhancer can result in reducing the viscosity of the cell culture, and/or increasing the extracellular concentration of the recombinant protein, and/or reducing the duration of cell culture to the point of harvest or protein recovery.

Abstract: The present invention generally relates to improved methods of assembly of two or more DNA fragments, methods of rapid ligation-independent cloning, and kits for rapid ligation-independent cloning and their uses.

Abstract: The present invention provides antibodies that bind to human interleukin-25 (IL-25) and methods of using the same. According to certain embodiments, the antibodies of the invention bind human IL-25 with high affinity. In certain embodiments, the invention includes antibodies that bind human IL-25 and block IL-25-mediated cell signaling. The antibodies of the invention may be fully human, non-naturally occurring antibodies. The antibodies of the invention are useful for the treatment of various disorders associated with IL-25 activity or expression, including asthma, allergy, chronic obstructive pulmonary disease (COPD), inflammatory bowel disease (IBD), including ulcerative colitis and Crohn's disease, atopic dermatitis (AD), and Eosinophilic Granulomatosis with Polyangiitis (EGPA), also know as Churg-Strauss Syndrome.

Abstract: The invention relates in one aspect to a pharmaceutical composition comprising a nucleic acid delivery vehicle for delivering a deliverable nucleic acid into a bacterial cell, wherein the delivery vehicle comprises a deliverable nucleic acid packaged into one or more bacteriophage coat proteins, and wherein the delivery vehicle is capable of infecting the bacterial cell to introduce the deliverable nucleic acid into the cell, following which the deliverable nucleic acid is capable of forming a plasmid in the cell and being transmitted to one or more different bacterial cells by conjugation and not by infection. Compositions including a pharmaceutical composition comprising the delivery vehicle, and methods involving use or manufacture of the delivery vehicle, are also disclosed.

Abstract: An antibody production process in mammalian cells in which engineered unpaired cysteine residues are post-translationally modified and capped with particular chemical entities, which capped antibodies are well suited to further site-specific conjugation steps to form antibody-drug conjugates (ADCs) or protein drug conjugates; ADCs produced using these capped antibodies including in particular ADCs formed by the selective reduction of the capped antibodies' cysteine residues, and ADCs formed using chemical handles such as aldehyde/azide/alkyne biorthogonal groups, which permit additional drug conjugation chemistry; and uncapped antibodies produced by cells in low cysteine, cysteine and glutathione media, and ADCs produced via direct conjugation to these uncapped antibodies.

Abstract: There is disclosed a process for in vitro synthesis and assembly of long, gene-length polynucleotides based upon assembly of multiple shorter oligonucleotides synthesized in situ on a microarray platform. Specifically, there is disclosed a process for in situ synthesis of oligonucleotide fragments on a solid phase microarray platform and subsequent, “on device” assembly of larger polynucleotides composed of a plurality of shorter oligonucleotide fragments.

Abstract: A mutant strain of the bacterium Clostridium botulinum having an inactivated botulinal neurotoxin gene is disclosed. The mutant strain contains an artificially created and inserted modified intron vector between nucleotides 580 and 581 of the sense strand of the gene. The mutant strain can be used in microbiological challenge testing of foods and food processing methods.

Abstract: This application is directed generally to minicircle DNA vectors for the vaccination of foot-and-mouth disease (FMD). The transgene expression cassette in the minicircle DNA vector includes: a eukaryotic translation initiation nucleotide sequence, a mutant nucleotide sequence that encodes a foot-and-mouth disease virus (FMDV) capsid polyprotein precursor that contains at least one mutation to eliminate a restriction enzyme recognition site, a nucleotide sequence that encodes a protease that cleaves the FMDV capsid polyprotein precursor into a plurality of FMDV capsid proteins and a translational regulatory element to regulate the expression of the protease. The minicircle DNA vectors can be transfected directly into the cell of a mammalian host. When transfected into the mammalian host cell, virus-like particles can be produced intrinsically to stimulate the mammalian host's immune system to develop adaptive immunity toward foot-and-mouth disease.

Abstract: The present invention provides minicircle nucleic acid vector formulations for use in administering to a subject, wherein the minicircle nucleic acid vectors include a polynucleotide of interest, a product hybrid sequence of a unidirectional site-specific recombinase, and are devoid of plasmid backbone bacterial DNA sequences. Also provided are methods of producing the subject formulations as well as methods for administering the minicircle nucleic acid vector formulations to a subject. The subject methods and compositions find use in a variety of different applications, including both research and therapeutic applications.

Abstract: Provided is a separatome-based recombinant peptide, polypeptide, and protein expression and purification platform based on the juxtaposition of the binding properties of host cell genomic peptides, polypeptides, and proteins with the characteristics and location of the corresponding genes on the host cell chromosome, such as that of E. coli, yeast, Bacillus subtilis or other prokaryotes, insect cells, mammalian cells, etc. This platform quantitatively describes and identifies priority deletions, modifications, or inhibitions of certain gene products to increase chromatographic separation efficiency, defined as an increase in column capacity, column selectivity, or both, with emphasis on the former. Moreover, the platform provides a computerized knowledge tool that, given separatome data and a target recombinant peptide, polypeptide, or protein, intuitively suggests strategies leading to efficient product purification.

Abstract: The present invention relates to methods for expressing nucleic acid sequences in prokaryotic host cells, where at least one DNA construct which is capable of episomal replication in a host cell and which comprises a nucleic acid sequence to be expressed under the transcriptional control of an L-rhamnose-inducible promoter, where the promoter is heterologous with regard to the nucleic acid sequence, is introduced into the host cell and the expression of he nucleic acid sequence is induced by addition of L-rhamnose, wherein the prokaryotic host cell is at least deficient with regard to an L-rhamnose isomerase.

Abstract: Nucleic acid molecules from Cannabis sativa (cannabis, hemp, marijuana) have been isolated and characterized, and encode polypeptides having aromatic prenyltransferase activity. Specifically, the enzyme, CsPT1, is a geranylpyrophosphate olivetolate geranyltransferase, active in the cannabinoid biosynthesis step of prenylation of olivetolic acid to form cannabigerolic acid (CBGA). Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds.

Abstract: The present invention encompasses a novel S. aureus bioconjugate vaccine. More generally, the invention is directed to Gram-positive and other bioconjugate vaccines containing a protein carrier, at least one polysaccharide such as a capsular Gram-positive polysaccharide, and, optionally, an adjuvant or pharmaceutically acceptable carrier. The instant invention also includes methods of producing Gram-positive and other bioconjugate vaccines. An N-glycosylated protein is also provided that contains one or more polysaccharides such as Gram-positive polysaccharides. The invention is additionally directed to engineered prokaryotic organisms comprising nucleotide sequences encoding a glycosyltransferase of a first prokaryotic organism and a glycosyltransferase of a second prokaryotic organism. The invention further includes plasmids and prokaryotic cells transformed with plasmids encoding polysaccharides and enzymes which produce an N-glycosylated protein and/or bioconjugate vaccine.

Abstract: The present disclosure provides a method for producing a Dicer polypeptide in a prokaryotic host cell. The present disclosure further provides a purified Dicer complex. The present disclosure further provides kits for producing a Dicer polypeptide in a prokaryotic host cell.

Abstract: The present invention pertains to vectors for regulating gene expression having at least one gene expressing cassette and at least one gene suppressing cassette, wherein the gene expression cassette encodes a polypeptide of interest, and wherein the gene suppressing cassette encodes a short interfering RNA (siRNA) molecule that reduces expression of a target gene by RNA interference. The present invention further includes vectors that contain suppressor cassettes in conjunction with cassettes upregulating gene expression regulated by either a constitutive promoter, such as a general CMV promoter, or a tissue specific promoter. The present invention further includes vectors that contain Dengue virus gene suppression cassettes. The present invention further includes pharmaceutical compositions containing such vectors, methods of modulating the expression of genes in a host using such vectors, and method of producing such vectors.

Abstract: The present disclosure provides temperature sensitive essential nucleic acid molecules from a psychrophilic bacterium, proteins encoded by the nucleic acid molecules, as well as recombinant cells into which have been introduced such nucleic acid molecules. The disclosed recombinant cells containing one or more essential nucleic acid molecules from a psychrophilic bacterium are thereby made temperature sensitive, and can be administered to a mammal to induce an immune response in the mammal.

Abstract: The present invention relates to recombinant N-glycosylated proteins, comprising one or more introduced N-glycosylated optimized amino acid sequence(s), nucleic acids encoding these proteins as well as corresponding vectors and host cells. In addition, the present invention is directed to the use of said proteins, nucleic acids, vectors and host cells for preparing medicaments. Furthermore, the present invention provides methods for producing said proteins.

Abstract: The present invention relates to a recombinant DNA expression/secretion system in E. coli wherein the said system combines the potential of signal peptide-based translocation of recombinant proteins to the periplasmic space of E. coli with membrane brave defective mutants of E. coli to further aid secretion into the extracellular space. The present invention further relates to the expression system which furthermore includes a helper plasmid to drive the expression of translocons to facilitate improved periplasmic secretion of the over-expressed recombinant protein. In addition, this system also facilitates efficient production of specific proteins of interest in E. coli.

Abstract: Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal lysyl-tRNAs, orthogonal lysyl-aminoacyl-tRNA synthetases, and orthogonal pairs of lysyl-tRNAs/synthetases, which incorporate homoglutamines into proteins are provided in response to a four base codon. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with homoglutamines using these orthogonal pairs.

Abstract: Nucleic acid molecules from hop (Humulus lupulus) have been isolated and characterized wherein said nucleic acid molecules encode polypeptides having aromatic prenyltransferase activity Expression or over-expression of said nucleic acid molecules alters the level of terpenophenolic compounds The polypeptides may be used in vivo or in vitro to produce terpenophenolic compounds (e g, prenylated acylphloroglucmols and prenylflavonoids) such as prenyl-PIVP, prenyl-PIBP, humulone, lupulone, desmethylxanthohumol and xanthohumol.

Abstract: The present invention relates to a method to engineer either the genome of a genetically modified organism, other bioengineered reagent, or in vitro translation system for protein synthesis from specific protein-coding genes so that the protein-coding genes so engineered can only produce proteins with an intended structure when translated within the context of that specifically engineered GMO or in vitro translation system. It also relates to nucleic acids for use in such GMOs or translation systems.

Abstract: Novel antimicrobial agents that can serve as replacements to conventional pharmaceutical antibiotics are disclosed. The antimicrobial agents comprise conjugatively transmissible plasmids that kill targeted pathogenic bacteria, but are not harmful to donor bacteria. Two types of lethal transmissible plasmids are disclosed. One type kills recipient bacteria by unchecked (“runaway”) replication in the recipient cells and is prevented from occurring in donor cells. Another type kills recipient bacteria by expressing a gene that produces a product detrimental or lethal to recipient bacterial cells, that gene being prevented from expression in donor cells.

Abstract: The present invention provides novel methods and materials for increasing the expression of recombinant polypeptides. Methods and materials of the invention allow increased expression of transcription units that include recombinant DNA sequences which encode polypeptides of interest. The present invention provides expression vectors which contain multiple copies of a transcription unit encoding a polypeptide of interest separated by at least one selective marker gene and methods for sequentially transforming or transfecting host cells with expression vectors to increase transcription unit dosage and expression.

Abstract: A method is disclosed for restoring a Glu+ phenotype to a PTS?/Glu? bacterial cell which was originally capable of utilizing a phosphotransferase transport system (PTS) for carbohydrate transport. Bacterial cells comprising the Glu+ phenotype have modified endogenous chromosomal regulatory regions which are operably linked to polynucleotides encoding galactose permeases and glucokinases.

Abstract: The invention provides molecular switches which couple external signals to functionality, and combinatorial methods of making and using the same involving circular permutation of nucleic acid and amino acid sequences. The switches according to the invention can be used, for example, to regulate gene transcription, target drug delivery to specific cells, transport drugs intracellularly, control drug release, provide conditionally active proteins, perform metabolic engineering, and modulate cell signaling pathways. Libraries comprising the switches, expression vectors and host cells for expressing the switches are also provided.

Abstract: Microorganisms are genetically engineered to synthesize caffeic acid from simple carbon sources via a tyrosine intermediate by means of a dual pathway that utilizes both endogenous and engineered enzymatic activities.

Abstract: The present invention is related to integrated method and tools to construct recombinant DNA molecules (to be used as DNA vaccine or gene therapy) without requiring the use of antibiotic(s) resistance gene(s) and without requiring the addition of one or more antibiotic(s) to the culture medium of cells submitted to this recombinant DNA method. The present invention allows to obtain the selection of recombinant host cell(s) transformed by a (exogenous) nucleic acid sequence of interest (extra-chromosomal vector containing the insert) and simultaneously stabilization (stable inheritance) of this (exogenous) nucleic acid sequence of interest into the transformed host cell(s) descendants (maintenance of the nucleic acid sequence of interest in the host cells population).

Abstract: Alterations utilizing nanoparticles. Certain embodiments of the invention are methods of delivering a substance to a target using a delivery-aid which includes nanoparticles. Those nanoparticles may be nanocarbon particles. Other embodiments are methods of delivering nanoparticles to a target involving placing a mask between a source of ballistic delivery of nanoparticles and the target. Other embodiments include irradiating a target to cause localized heating of the region of the target in which the nanodiamonds or OLC particles are present. Other embodiments utilize nanoparticles to make cells competent for genetic transformation. This abstract is not to be considered limiting, since other embodiments may deviate from the features described in this abstract.

Abstract: The present invention provides a novel method of producing a recombinant bacterium for production of a non-natural protein, including: (1) expressing tRNA in a bacterium, which tRNA recognizes UAG codon; (2) expressing an aminoacyl-tRNA synthetase in the bacterium, which aminoacyl-tRNA synthetase acylates the tRNA with a non-natural amino acid or an ?-hydroxy acid; (3) (i) introducing a DNA construct into the bacterium, which DNA construct is for expressing, in the absence of a release factor for terminating translation at UAG codon, a function of at least one gene selected from the group consisting of genes each of which loses its function when a gene that codes for the release factor is defective and/or introducing an alteration into said at least one gene in a chromosome of the bacterium, which alteration is for expressing the function of said at least one gene in the absence of the release factor; and (4) causing the gene that codes for the release factor in the bacterium to be defective.

Abstract: Novel antimicrobial agents that can serve as replacements to conventional pharmaceutical antibiotics are disclosed. The antimicrobial agents comprise conjugatively transmissible plasmids that kill targeted pathogenic bacteria, but are not harmful to donor bacteria. Two types of lethal transmissible plasmids are disclosed. One type kills recipient bacteria by unchecked (“runaway”) replication in the recipient cells and is prevented from occurring in donor cells. Another type kills recipient bacteria by expressing a gene that produces a product detrimental or lethal to recipient bacterial cells, that gene being prevented from expression in donor cells.

Abstract: This invention provides methods and compositions for incorporation of an unnatural amino acid into a peptide using an orthogonal aminoacyl tRNA synthetase/tRNA pair. In particular, an orthogonal pair is provided to incorporate 5-hydroxy-L-tryptophan in a position encoded by an opal mutation.

Abstract: The invention relates to orthogonal pairs of tRNAs and aminoacyl-tRNA synthetases that can incorporate the unnatural amino acid phenylselenocysteine into proteins produced in eubacterial host cells such as E. coli. The invention provides, for example but not limited to, novel orthogonal aminoacyl-tRNA synthetases, polynucleotides encoding the novel synthetase molecules, methods for identifying and making the novel synthetases, methods for producing proteins containing the unnatural amino acid phenylselenocysteine and translation systems. The invention further provides methods for producing modified proteins (e.g., lipidated proteins) through targeted modification of the phenylselenocysteine residue in a protein.

Abstract: The present invention is directed generally to compositions and methods for obtaining secretion of antibodies or antigen-binding antibody fragments from prokaryotes without the need for a signal peptide through making use of mutant host strains with altered secretory properties. In particular, the invention provides host cells and methods for obtaining secretion of antibodies or antigen-binding antibody fragments from bacteria without the need for a signal peptide and provides diverse libraries of antibody sequence resulting from such methods. The invention additionally provides diverse libraries.

Abstract: The invention provides microbial strains possessing improved properties for production of aspartate-derived amino acids and chemicals. Methods of making such strains are provided. These methods include altering expression of the aceBAK operon, the glcB gene, or both. Alteration of expression may be accomplished through increased transcription, relief from native transcriptional control, and/or other means. Replacement of native promoters for these genes is also contemplated; for instance, their native promoters may be replaced by the tac promoter (Ptac).

Abstract: Escherichia coli strains, such as enterotoxigenic E. coli strains, genetically engineered to express from recombinant plasmids one or more colonization factors (CFs) associated with enterotoxigenic Escherichia coli bacteria (ETEC) in an increased amount compared to said CFs expressed by ETEC wild-type reference strains, as well as a method of producing such strains, and vaccine compositions against diarrhea comprising such strains, are described. Further, E. coli strains expressing unnatural combination of at least two different CFs, e.g., CFA/I+CS2, CFA/I+CS6, or CS2+CS4 are disclosed.

Abstract: The instant invention provides methods and compositions for generating recombinant adenoviral vectors. The invention also provides kits comprising for the generation of recombinant adenoviral vectors.

Abstract: The present invention relates to a mutant bacterial PRPP synthetase which is resistant to feedback by purine nucleotides, and a method for producing L-histidine using the bacterium of the Enterobacteriaceae family wherein the L-amino acid productivity of said bacterium is enhanced by use of the PRPP synthetase which is resistant to feedback by purine nucleotides, coded by the mutant prsA gene.

Abstract: The present invention features homologous recombination methods and systems. The methods and systems promote highly efficient homologous recombination in cells (e.g., in prokaryotic cells). The methods and systems are useful, for example, in pharmaceutical drug development, vaccine development and cloning.

Abstract: Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal lysyl-tRNAs, orthogonal lysyl-aminoacyl-tRNA synthetases, and orthogonal pairs of lysyl-tRNAs/synthetases, which incorporate homoglutamines into proteins are provided in response to a four base codon. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with homoglutamines using these orthogonal pairs.

Abstract: Disclosed are a recombinant expression vector comprising a nucleotide sequence encoding an E. coli-derived signal sequence and a nucleotide sequence encoding an immunoglobulin constant region, and a transformant transformed with the expression vector. Also, disclosed is a method of mass-producing an immunoglobulin constant region by culturing the transformant and expressing the immunoglobulin constant region in a water-soluble form.

Abstract: Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate redox active amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with redox active amino acids using these orthogonal pairs.

Abstract: The invention provides methods and nucleic acid constructs to express clostripain. The source of the coding region for recombinantly expressed clostripain is Clostridium histolyticum.

Abstract: Methods of increasing yields of succinate using aerobic culture methods and a multi-mutant E. coli strain are provided. Also provided is a mutant strain of E. coli that produces high amounts of succinic acid.

Abstract: The present invention provides a method for producing an L-amino acid using a bacterium of the Enterobacteriaceae family, particularly a bacterium belonging to genus Escherichia or Pantoea, which has been modified to attenuate expression of the cpxR gene.

Abstract: The invention provides methods and compositions for in vivo incorporation of unnatural amino acids. Also provided are compositions including proteins with unnatural amino acids.

Abstract: The present invention provides an agent that modulates physiological condition of pests, wherein the agent has an ability to modulate the activity of an insect choline acetyltransferase; a method for assaying pesticidal activity of a test substance, which comprises measuring the activity of a choline acetyltransferase in a reaction system in which the choline acetyltransferase contacts with a test substance, and the like.

Abstract: An object is to provide a means of highly producing an oxalate decarboxylase originating in a microorganism. A recombinant expression plasmid vector, which contains an ?-amylase promoter belonging to the genus Bacillus and an oxalate decarboxylase gene originating in a microorganism that is provided under the control of the promoter, is constructed. A host bacterium is transformed with this vector to prepare an oxalate decarboxylase producing bacterium. A recombinant oxalate decarboxylase is produced by culturing the producing bacterium and then recovering the oxalate decarboxylase thus produced.

Abstract: Provided is a method of producing a template DNA used for protein synthesis comprising a step of amplifying a linear double-stranded DNA by polymerase chain reaction (PCR), by using a reaction solution comprising a first double-stranded DNA fragment comprising a sequence coding for a protein or a portion thereof, a second double-stranded DNA fragment comprising a sequence overlapping with the 5? terminal region of the first DNA fragment, a third double-stranded DNA fragment comprising a sequence overlapping with the 3? terminal region of the first DNA fragment, a sense primer which anneals with the 5? terminal region of the second DNA fragment, and an anti-sense primer which anneals with the 3? terminal region of the third DNA fragment, wherein the second DNA fragment comprises a regulatory sequence for transcription and translation of a gene, and the concentrations of the second DNA fragment and the third DNA fragment in the reaction solution each range from 5 to 2,500 pmol/L.