Abstract: The present invention relates to a method based on the use of restriction enzyme digestion and ligation via cleavage sites, thereby to prepare two or more standardized expression cassettes.

Abstract: The present invention relates to a method for preparing a mutant E. coli strain, capable of simultaneously using glucose and xylose, by genetic engineering and evolutionary adaptation; the mutant E. coli prepared using the same; and a method for producing biofuels, biologically active ingredients, medicinal materials or base chemicals for the chemical industry using the mutant E. coli. Being capable of simultaneously using glucose and xylose, in contrast to wild-type E. coli, the mutant E. coli can be effectively applied to the enzymatic saccharification process of producing biofuels from a biomass.

Abstract: The present invention discloses a method for introducing an exogenous DNA by overcoming the restriction modification barrier of the target bacterium. The method provided in the present invention comprises the steps of 1) co-expressing all DNA-methyltransferase-encoding genes in the genome of the target bacterium in E. coli in which the restriction modification system thereof has been deleted to obtain a recombinant bacterium A; 2) introducing an exogenous DNA molecule into the recombinant bacterium A for in vivo modification so as to obtain a methylation-modified exogenous DNA molecule; 3) introducing the methylation-modified exogenous DNA molecule into the target bacterium. The experiments of the invention have demonstrated that the invention has a high transformation efficiency compared to prior methods for enabling genetic manipulation by overcoming the restriction modification barrier of the bacterium.

Abstract: The present invention relates to a polynucleotide encoding an enzyme having carboxyl esterase [E.C. 2.1.1.1] activity.

Abstract: A yeast cell comprising LDH from a Sordaria genus fungi, in which activity of lactate dehydrogenase converting pyruvate into lactate is increased, as well as a method of preparing the yeast cell and a method of using the yeast cell to produce lactate.

Abstract: An inactive CRISPR/Cas system-based fusion protein and its applications in gene editing are disclosed. More particularly, chimeric fusion proteins including an inCas fused to a DNA modifying enzyme and methods of using the chimeric fusion proteins in gene editing are disclosed. The methods can be used to induce double-strand breaks and single-strand nicks in target DNAs, to generate gene disruptions, deletions, point mutations, gene replacements, insertions, inversions and other modifications of a genomic DNA within cells and organisms.

Abstract: A recombinant Corynebacterium genus microorganism, and a method of producing C4 dicarboxylic acid under anaerobic conditions using the Corynebacterium genus microorganism.

Abstract: The present invention relates to a method for producing 3-hydroxypropionic acid by culturing in a glycerol-containing medium a mutant microorganism obtained by inserting or amplifying a gene encoding propanediol utilization protein in a microorganism having the abilities to produce coenzyme B12 and produce 3-hydroxypropionic acid using glycerol as a carbon source. According to the present invention, 3-hydroxypropionic acid can be produced in high yield from glycerol without having to add expensive coenzyme B12 as a cofactor.

Abstract: Site-specific Listeria integration vectors and methods for their use are provided. The subject vectors include a bacteriophage integrase gene and a bacteriophage attachment site, where in many embodiments the bacteriophage that is the source 0 of these elements is a listeriophage. In certain embodiments, the subject vectors further include a multiple cloning site, where the multiple cloning site may further include a polypeptide coding sequence, e.g., for a heterologous antigen. The subject vectors and methods find use in a variety of different applications, including the study of Listeria species and the preparation of Listeria vaccines.

Abstract: Recombinant microbial cells and methods for producing 5-hydroxytryptophan (5HTP) using such cells are described. More specifically, the recombinant microbial cell comprises an exogenous gene encoding an L-tryptophan hydroxylase, and means for providing tetrahydrobiopterin (THB). Related sequences and vectors for use in preparing such recombinant microbial cells are also described.

Abstract: The present invention relates to a method for carrying out recombination at a target locus.

Abstract: The present invention relates to a method for producing a product of a reaction catalysed by a nitrilase, which method comprises the steps (i) providing a microorganism comprising said nitrilase located on its surface, and/or a membrane preparation of said microorganism, and (ii) contacting the microorganism and/or the membrane preparation thereof with one or more nitrilase substrates under conditions compatible with nitrilase activity. The present invention further relates to a method for producing enantiomerically pure (R)-mandelic acid using the nitrilase-displaying whole cell biocatalyst or membrane preparation thereof for the conversion of racemic mandelonitrile.

Abstract: Establishment of an effective and uniform vaccine development strategy is key to conquering current and emerging infectious diseases. Despite successes against an array of bacterial agents, current approaches to vaccine development are as diverse as the microbes they target and require adjuvants that often have limited efficacy and/or toxic side effects. As a consequence, vaccine discovery is often slow, inefficient, and unsuccessful in the case of many high priority pathogens. The present disclosure suggests that vaccine generation for bacterial pathogens can be improved by optimizing the efficiency of processing/presentation of a bacterial immunogen via the targeting of immunogen to CR2 and/or TLR2 on APCs. This approach not only yields an adjuvant-free mucosal vaccine against a Category A biothreat agent, but also establishes a novel genetic approach/platform for vaccine development, which is applicable to many other infectious agents, thereby profoundly impacting preventive medicine/public health.

Abstract: The invention provides for systems, methods, and compositions for manipulation of sequences and/or activities of target sequences. Provided are vectors and vector systems, some of which encode one or more components of a CRISPR complex, as well as methods for the design and use of such vectors. Also provided are methods of directing CRISPR complex formation in eukaryotic cells and methods for selecting specific cells by introducing precise mutations utilizing the CRISPR/Cas system.

Abstract: It is an object of the present invention to provide a substance usable as an anticancer agent or DDS, which has intracellular stability, which is capable of evading side effects from functional disorder with respect to normal cells, or which has instantaneous effect. The inventors developed a novel chimeric peptide targeting cancer cells which overexpress EGFR or the like using a binding peptide such as a peptide sequence binding to EGFR, and a lytic peptide sequence, thereby solving such an object. Particularly, by using a chimeric peptide including an EGF receptor-binding peptide or the like and a cytotoxic peptide, this object was solved.

Abstract: It is an object of the present invention to provide a substance usable as an anticancer agent or DDS, which has intracellular stability, which is capable of evading side effects from functional disorder with respect to normal cells, or which has instantaneous effect. The inventors developed a novel chimeric peptide targeting cancer cells which overexpress EGFR or the like using a binding peptide such as a peptide sequence binding to EGFR, and a lytic peptide sequence, thereby solving such an object. Particularly, by using a chimeric peptide including an EGF receptor-binding peptide or the like and a cytotoxic peptide, this object was solved.

Abstract: The invention provides an isolated genetically modified non-mammalian organism, wherein the activity of acyl-CoA:sterol acyltransferase/sterol O-acyltransferase (EC 2.3.1.26) and/or diacylglycerol acyltransferase/diacylglycerol O-acyltranferase (EC 2.3.1.20) and/or lecithin cholesterol acyl transferase/phospholipid: diacylglycerol acyltransferase (EC 2.3.1.158) and/or acyl CoA-wax alcohol acyltransferase (EC 2.3.1.75) is reduced or abolished in comparison with a corresponding wildtype organism, methods of use of such an organism, shuttle vehicles for making such an organism and methods for producing such an organism.

Abstract: Provided are fusion proteins of diacylglycerol acyltransferase (DGAT) polypeptides fused to at least one heterologous intracellular localization domain, such as a bacterial membrane-targeting domain, modified photosynthetic microorganism that comprise such fusion proteins, and related methods for improving the production of lipids by photosynthetic microorganisms, including lipids such as triglycerides, fatty acids, and wax esters.

Abstract: To produce a bacterial microcompartment shell, or a designed shell based on naturally occurring bacterial microcompartment shells in a new host organism, a synthetic operon is constructed that contains the desired shell protein genes and translation efficiency is controlled by host specific ribosomal binding sites. Proteins or other molecules can be encapsulated in the microcompartment shells by various methods described herein. The constructs can also be used to express self-assembling sheets comprised of shell proteins.

Abstract: Recombinant microbial cells and methods for producing melatonin and related compounds using such cells are described. More specifically, the recombinant microbial cell may comprise exogenous genes encoding one or more of an L-tryptophan hydroxylase, a 5-hydroxy-L-tryptophan decarboxylyase, a serotonin acetyltransferase, an acetylserotonin O-methyltransferase; an L-tryptophan decarboxy-lyase, and a tryptamine-5-hydroxylase, and means for providing tetrahydrobiopterin (THB). Related sequences and vectors for use in preparing such recombinant microbial cells are also described.

Abstract: The present disclosure relates to recombinant host cells comprising one or more recombinant polynucleotides encoding enzymes in select pathways that provide the ability to use the cells to produce 1,3-butadiene. The present disclosure also provides methods of manufacturing the recombinant host cells, and methods for the use of the cells to produce 1,3-butadiene. The methods utilize recombinant host cells that comprise an engineered pathway of enzymes that provides for the conversion of naturally occurring intermediate crotonyl-CoA (or -ACP) to 1,3-butadiene through enzyme catalyzed steps involving the reduction of glutaconyl-CoA (or -ACP) to form the intermediate 5-hydroxypent-3-enoate. The disclosure provides alternative engineered pathway involving either decarboxylation of 5-hydroxypent-3-enoate directly to 1,3-butadiene, or phosphorylation of 5-hydroxypent-3-enoate followed by a phosphate elimination step catalyzed by a decarboxylase to produce 1,3-butadiene.

Abstract: The present invention provides compositions and methods for targeted cleavage of cellular chromatin in a region of interest and/or homologous recombination at a predetermined site in cells. Compositions include fusion polypeptides comprising a TAL effector binding or a zinc finger domain and an I-TevI homing endonuclease cleavage domain as well as nucleic acid sequence encoding the same. The use of the I-TevI domain allows for monomer endonuclease sequences to achieve cleavage of cellular chromatin and represents an advantage over prior endonucleases which require self-dimerization, and two nucleases with appropriate spacers.

Abstract: A method for carrying out recombination at a target locus in a Rasamsonia cell, which method comprises: providing two or more nucleic acids which, when taken together, comprise: (a) sequences capable of homologous recombination with sequences flanking the target locus; (b) two or more site-specific recombination sites; (c) a sequence encoding a recombinase which recognizes the site-specific recombination sites; and (d) a sequence encoding a marker, wherein the two or more nucleic acids are capable of homologous recombination with each other so as to give rise to a single nucleic acid, and wherein at least two of the two or more nucleic acids each comprise a sequence encoding a non-functional portion of the marker; and recombining in a Rasamsonia cell the said two or more nucleic acids with each other and with the sequences flanking the target locus so that a contiguous nucleic acid sequence encoding a functional marker and the sequence encoding the recombinase are inserted at the target locus, said marker-enc

Abstract: The invention provides methods and nucleic acid constructs that may be used to modify a nucleic acid of interest at a target locus within the genome of a host. In some aspects, the invention contemplates producing in vivo a gene targeting substrate (GTS), which may be comprised of both DNA and RNA components. The gene targeting substrate may comprise a gene targeting nucleotide sequence (GTNS), which is homologous to the target locus, but comprises a sequence modification compared to the target locus. The gene targeting substrate may be produced by reverse transcription of a gene targeting message RNA (gtmRNA). The gene targeting message RNA may be folded for self-priming for reverse transcription by a reverse transcriptase. The gene targeting message RNA may in turn be the product of transcription of a gene targeting construct (GTC) encoding the gene targeting message RNA.

Abstract: The present invention relates to a transformation vector comprising the partial fragments of a gene encoding transposase, a microorganism transformed with the vector, and a method of producing lysine using the microorganism.

Abstract: A thermophilic microorganism is modified to permit the increased production of ethanol, wherein a first modification is the inactivation of the lactate dehydrogenase gene and a second modification upregulates the pyruvate dehydrogenase gene.

Abstract: Methods and constructs are provided for controlling processes in live animals, plants or microbes via genetically engineered near-infrared light-activated or light-inactivated proteins including chimeras including the photosensory modules of bacteriohytochromes and output modules that possess enzymatic activity and/or ability to bind to DNA, RNA, protein, or small molecules. DNA encoding these proteins are introduced as genes into live animals, plants or microbes, where their activities can be turned on by near-infrared light, controlled by the intensity of light, and turned off by near-infrared light of a different wavelength than the activating light. These proteins can regulate diverse cellular processes with high spatial and temporal precision, in a nontoxic manner, often using external light sources.

Abstract: A method for expression of engineered constructs containing sequences coding for functional genes of interest in E. coli strains. Here tceA and tceB are fused to an inducible, active promoter to optimize transcription of the open reading frames, and an introduced consensus Shine-Dalgarno sequence allowing for optimal ribosome binding and translation of the open reading frames, and codons optimized for expression in E. coil. This over-expressed protein is then used to design appropriate mass spectro metric methods for environmental detection.

Abstract: Provided herein are nucleic acid constructs that contain a synthetic control element that includes a cis-regulator of translation, and an adapter translation-coupled regulator of transcription. Further provided herein are nucleic acid constructs that contain nucleic acid sequences under the control of the synthetic control elements. Also provided are compositions and methods related to the nucleic acid constructs.

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 invention relates to fungal strains having at least one genetic modification which leads to a reduction of the activity of at least one fungal carboxylic acid transporter and to a method for producing or using said fungal strains.

Abstract: The invention relates to the production of one or more terpenoids through microbial engineering, and relates to the manufacture of products comprising terpenoids.

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: Provided herein are recombinant cells (e.g., recombinant bacteria or plant, insect, mammalian, and yeast cells) containing a nucleic acid encoding a CYP97A protein or a nucleic acid encoding a CYP97B protein; a nucleic acid encoding a CYP97C protein; a nucleic acid encoding a geranylgeranyl pyrophosphate synthase protein; a nucleic acid encoding a phytoene synthase protein; a nucleic acid encoding a phytoene desaturase protein; a nucleic acid encoding a lycopene ?-cyclase protein; and a nucleic acid encoding a lycopene ?-cyclase protein. Also provided are methods of producing lutein that include culturing these recombinant cells (e.g., recombinant bacteria and yeast cells), and methods of generating these recombinant cells (e.g., recombinant bacteria and yeast cells). Also provided is lutein produced by these methods, and pharmaceutical compositions, food supplements, food products, and cosmetic compositions that contain lutein produced by these methods.

Abstract: The present invention relates to isolated polypeptides having cellulolytic enhancing activity and isolated polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The present invention provides methods and compositions for increasing oil content in algae. More particularly, the present invention relates to enhancement of phytohormone activity in algae, by genetic transformation thereof or by supplementing the algal growth medium with phytohormones, to maximize the production of oil within the algae cells.

Abstract: Various aspects and embodiments of the present disclosure are directed to methods and compositions for functionalizing endogenous bacteria in vivo. The methods include delivering to endogenous bacterial cells a recombinant bacteriophage or phagemid that is engineered to contain at least one genetic circuit.

Abstract: The present invention refers to single-chain fusion proteins comprising three soluble TNF superfamily (TNFSF) cytokine domains and nucleic acid molecules encoding these fusion proteins. The fusion proteins are substantially non-aggregating and suitable for therapeutic, diagnostic and/or research applications.

Abstract: A modified microorganism for producing a high yield of 1,4-butanediol, and a method of producing 1,4-butanediol using the modified microorganism, wherein the modified microorganism is altered so as to delete or disrupt expression of at least one of transcription regulatory factor NCg12886, GCN5-related N-acetyltransferase NCg12090, hypothetical protein NCg10224 or sugar phosphate isomerase/epimerase NCg12956.

Abstract: The present invention provides reagents and methods for improved homologous recombination.

Abstract: The present invention provides yeast having increased resistance to freezing stress in which POG1 gene is inactivated, a method for producing such yeast, and a method for using such yeast in food production.

Abstract: The present invention concerns micro-organisms which present cellulases on their surface. Corresponding micro-organisms were produced with the aid of corresponding plasmids which encode a section comprising a signal peptide, a heterologous cellulase, an optional protease recognition site, a transmembrane linker and a transporter domain of an autotransporter or a variant thereof. Such micro-organisms were advantageously used in the conversion of cellulose into cellobiose and/or glucose. It was also possible to recover the micro-organisms from the reaction mixture following conversion from simple substrates. Also, a combination of various micro-organisms, which were populated with exocellulases, endocellulases and beta-glucosidases, were used to produce glucose from cellulose or wood.

Abstract: The present invention relates to microorganisms and polypeptides for detoxifying aldehydes associated with industrial fermentations. In particular, a heat-stable, NADPH- and iron-dependent alcohol dehydrogenase was cloned from Thermoanaerobacter pseudethanolicus 39E and displayed activity against a number of aldehydes including inhibitory compounds that are produced during the dilute-acid pretreatment process of lignocellulosic biomass before fermentation to biofuels. Methods to use the microorganisms and polypeptides of the invention for improved conversion of bio mass to biofuel are provided as well as use of the enzyme in metabolic engineering strategies for producing longer-chain alcohols from sugars using thermophilic, fermentative microorganisms.

Abstract: A filamentous fungal cell having reduced UDP-galactofuranose is provided. The fungal cell may, in certain embodiments, contain a nuclear genome comprising an inactivated UDP-galactopyranose mutase (UDP-galp mutase) gene and a recombinant nucleic acid for expression of a protein. Also provided are methods of producing a protein using the subject fungal cell, as well as methods of producing the subject fungal cell.

Abstract: A mutant Escherichia coli ?-barrel monomeric protein pore bioengineered to remove a lumen-occluding domain and modify some of its extracellular domains. The modified protein, FhuA ?/C?4L, forms a highly conductive pore as compared to other known pores and is capable of sensing large polypeptides and discriminating between modified protein analytes.

Abstract: Disclosed herein are transgenic cells expressing a heterologous nucleic acid encoding a prephenate dehydrogenase (PDH) protein, a heterologous nucleic acid encoding a homogentisate solanesyl transferase (HST) protein, a heterologous nucleic acid encoding a deoxyxylulose phosphate synthase (DXS) protein, or a combination of two or more thereof. In particular examples, the disclosed transgenic cells have increased plastoquinone levels. Also disclosed are methods of increasing cell growth rates or production of biomass by cultivating transgenic cells expressing a heterologous nucleic acid encoding a PDH protein, a heterologous nucleic acid encoding an HST protein, a heterologous nucleic acid encoding a DXS protein, or a combination of two or more thereof under conditions sufficient to produce cell growth or biomass.

Abstract: The present invention relates to inducing acid resistance in a bacterium and methods of increasing the acid resistance of an acid sensitive bacterium.

Abstract: The present invention relates to recombinant cells, particularly recombinant plant cells, which are capable of producing dihydrosterculic acid and/or derivatives thereof. The present invention also relates to methods of producing oil comprising dihydrosterculic acid and/or derivatives thereof.

Abstract: Provided are isolated polypeptides having beta-xylosidase activity and polynucleotides encoding the polypeptides. Also provided are nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

Abstract: The endogenous pnp gene encoding polynucleotide phosphorylase in the Zymomonas genome was identified as a target for modification to provide improved xylose utilizing cells for ethanol production. The cells are in addition genetically modified to have increased expression of ribose-5-phosphate isomerase (RPI) activity, as compared to cells without this genetic modification, and are not limited in xylose isomerase activity in the absence of the pnp modification.