Abstract: The invention relates to a method for modifying a template double stranded polynucleotide, especially for characterisation using nanopore sequencing. The method produces from the template a plurality of modified double stranded polynucleotides. These modified polynucleotides can then be characterised.

Abstract: The invention relates to an isolated, genetically modified, living non-mammal organism, having increased HMG-CoA-reductase activity compared to the wild type, and having reduced C24-methyltransferase and/or delta22-desaturase activity compared to the wild type. The invention is characterized in that the organism has increased dehydrocholesterol-delta70-reductase activity compared to the wild type. The invention further relates to different uses of such an organism, to a test kit comprising such an organism, and to a membrane extract of such an organism.

Abstract: Systems and methods are described for parallel macromolecular delivery and biochemical/electrochemical interface to whole cells employing carbon nanostructures including nanofibers and nanotubes. A method includes providing a first material on at least a first portion of a first surface of a first tip of a first elongated carbon nanostructure; providing a second material on at least a second portion of a second surface of a second tip of a second elongated carbon nanostructure, the second elongated carbon nanostructure coupled to, and substantially parallel to, the first elongated carbon nanostructure; and penetrating a boundary of a biological sample with at least one member selected from the group consisting of the first tip and the second tip.

Abstract: The present provides selection markers, methods, nucleic acids, and vectors of use in the preparation of recombinant Clostridium spp.

Abstract: The invention refers to a method of producing a recombinant polypeptide of interest (POI) in a cell culture, comprising genetically engineering a eukaryotic cell line—to specifically cause prolongation of the G2+M cell cycle phase in a pre-culture phase, and—to produce the POI in a producing phase following the pre-culture phase, a high producer cell line and cell culture as well as a method of increasing the yield of a recombinant POI production in a cell culture.

Abstract: To provide a technology that solves the above-mentioned problems, and is directly applicable to eukaryotic algal cells with cell-wall, the technology enabling gene transfer and transformation to be performed with high efficiency and good reproducibility irrespective of species of algae.

Abstract: The present invention includes devices and methods for transfecting a cell or cell population and dynamic monitoring of cellular events. A variety of microelectronic devices are provide that incorporate functions such as electroporation, modulation of a transmembrane potential and dynamic monitoring of cellular functions and mechanisms.

Abstract: An object of the present invention is to provide a method of efficiently constructing a gene delivery carrier having a favorable activity and expression efficiency of a protein expressed by a gene introduced by transformation. Moreover, an object of the present invention is to provide a pharmaceutical composition comprising a gene delivery carrier constructed by the construction method and a therapeutic agent for solid tumor comprising the resistant bacterium.

Abstract: Methods of cloning insert sequences into cloning vectors with high efficiency and in the correct orientation are described. In one aspect, the invention features a method of producing a plasmid comprising an insert fragment and a vector fragment in a predetermined orientation. In some embodiments, the method includes cleaving a first nucleotide sequence at a plurality of sites with a first restriction enzyme to generate a first population of nucleotide fragments, the first population of nucleotide fragments comprising insert fragments and non-insert fragments, the insert fragments comprising a non-palindromic overhang at a 5? end, at a 3? end, or at both.

Abstract: The present invention is directed to particular monoclonal antibodies and fragments thereof that find use in the detection, prevention and treatment of influenza virus infections. In particular, these antibodies may neutralize or limit the replication of H1N1 influenza virus. Also disclosed are improved methods for producing such monoclonal antibodies.

Abstract: The present invention relates to methods and compositions for engineering sporulating bacterial cells, particularly a cell of the class Clostridia. In particular, the present invention relates to the generation of sporulation deficient bacteria for the generation of industrial superior phenotypes.

Abstract: The present invention relates to plasmid curing, and particularly to efficient and stress-free methods for displacing resident or endogenous plasmids from a host cell, such as a bacterium. The invention extends to method of displacing a plasmid comprising a post-segregational killing system from a host cell, the method comprising introducing a recombinant nucleic acid molecule into a host cell harboring a plasmid comprising a post-segregational killing (PSK) system, characterized in that the recombinant nucleic acid molecule is adapted to neutralize the toxic effects of the plasmid's post-segregational killing system, and wherein the nucleic acid molecule is also adapted to outcompete or inhibit replication of the plasmid. The invention further extends to recombinant nucleic acid molecules that can be used in this method, as well as further uses of the methods and nucleic acid molecules of the invention.

Abstract: The invention provides a composition useful to prepare influenza viruses, e.g., in the absence of helper virus, using vectors which include tandem transcription cassettes containing PolI and/or PolII promoters.

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 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: This invention provides transcription regulatory control sequences, the activity of which function as biomarkers for a variety of biological responses. This invention also provides expression constructs in which a biomarker transcription regulatory sequence is operably linked with a sequence for a reporter. Cells that comprise these expression constructs can be used in assays to identify conditions that modulate activity of the biological response.

Abstract: The present invention relates to a gene associated with ethanol tolerance, and yeast strains and uses using the same. The yeast strain of this invention may growth under the condition not only with high-concentration ethanol, preferably 6-15% ethanol, but also in high osmotic pressure, preferably 30-40% glucose or sucrose. The present inventors developed yeast strains resistant to high-concentration glucose and ethanol, suggesting that they would be valuably applied to much effective ethanol production, and also be utilized as a superbacteria having tolerance to various stresses for ethanol production with high efficiency.

Abstract: The present invention relates to the use of one or more cas genes for modulating resistance in a cell against a target nucleic acid or a transcription product thereof.

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: Zymomonas is unable to synthesize pantothenic acid and requires this essential vitamin in growth medium. Zymomonas strains transformed with an operon for expression of 2-dehydropantoate reductase and aspartate 1-decarboxylase were able to grow in medium lacking pantothenic acid. These strains may be used for ethanol production without pantothenic acid supplementation in seed culture and fermentation media.

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: This invention relates to binding members, especially antibody molecules, specific for interleukin 1 receptor 1 (IL-1R1). For example, isolated binding members specific for IL-1R1 which competes with IL-1 and IL-1Ra for binding to IL-1R1 and binds Il-1R1 with a KD of 10 pM or less when measured by Kinexa™. The binding members are useful for, inter alia, treatment of disorders mediated by IL-1R1 including rheumatoid arthritis, asthma and chronic obstructive pulmonary disease (COPD).

Abstract: The present invention relates to methods and compositions for engineering Clostridia species. In particular, embodiments of the present invention relate to the expression of recombinant resolvase proteins in Clostridia species.

Abstract: The technology relates in part to biological methods for producing adipic acid and engineered microorganisms capable of such production.

Abstract: Microalgae are potential energy resources for production of biofuels, such as biodiesel, ethanol, and butanol. A method for enhancing cell growth of microalgae enhances transgenic expression of a bicarbonate transporter (HCO3? transporter) in microalgae and thereby obtains a genetically modified microalgae capable of enhanced inorganic carbon fixation, efficient photosynthesis, and expeditious cell growth. The genetically modified microalgae are fit for use in biofuel production.

Abstract: The present invention provides novel C. botulinum conjugatively transmissible plasmids and methods of use thereof. Specifically, described herein are novel, conjugatively transmissible clostridial plasmids which are capable of being transferred among and between clostridial species. The novel plasmids of the present invention therefore permits the delivery of heterologous clostridial genes into a clostridial host, such as C. botulinum, and the expression of genes of interest in that host, including clostridial toxins and the nontoxigenic components of the toxin complex, toxin fragments, or antigenic portions thereof, in a way both that ensures abundant expression and facilitates purification. Furthermore, toxins with altered structures, chimeric, hybrid toxins, and other toxin derivatives valuable in medicine could be synthesized in this system.

Abstract: Xylose-utilizing Zymomonas strains studied were found to accumulate ribulose when grown in xylose-containing media. Engineering these strains to increase ribose-5-phosphate isomerase activity led to reduced ribulose accumulation, improved growth, improved xylose utilization, and increased ethanol production.

Abstract: The object is to provide a transformant which can produce a heterologous protein having a structurally controlled O-linked sugar chain having an O-Man-Gal disaccharide structure, a method for producing the transformant by using Schizosaccharomyces pombe as the host, and provide a host for producing the transformant and a method for producing an O-glycosylated heterologous protein. An Schizosaccharomyces pombe host having no omh1 gene or an inactivated omh1 gene in its chromosomes for producing an O-glycosylated heterologous protein having an O-linked sugar chain having an O-Man-Gal disaccharide structure by expression of the heterologous protein by a genetic engineering technique and subsequent glycosylation of the expressed heterologous protein. A transformant from the host, a method for producing the transformant and a method for producing an O-glycosylated heterologous protein by using the transformant.

Abstract: The subject invention pertains to the discovery that the NADH-dependent propanediol oxidoreductase (FucO) can reduce furfural. This allows for a new approach to improve furfural tolerance in bacterial and/or yeast cells used to produce desired products. Thus, novel biocatalysts (bacterial, fungal or yeast cells) exhibiting increased tolerance to furfural and 5-hydroxymethylfurfural (5-HMF) are provided as are methods of making and using such biocatalysts for the production of a desired product.

Abstract: A system for ligase-free cloning and/or expressing a target gene is described herein. A preferred version of the invention includes an E. coli host. The host preferably includes a T7 RNA polymerase gene comprising a T7gpl coding sequence, a lacUV5 promoter, and a lac operator. The host preferably further includes a lacI gene comprising a lacI coding sequence with an ATG start codon, a promoter derived from the lacql allele, and a translational enhancer derived from a 5? RNA leader sequence of T7 gene 10. The invention further includes a low-copy plasmid vector comprising a T7 promoter a lac operator operationally linked to the T7 promoter. The system is configured to inhibit target gene expression when uninduced and to permit gene expression upon induction by auto-induction.

Abstract: Provided are methods for introducing a molecule of interest into a plant cell having a cell wall by using a QD-peptide conjugate having a quantum dot (QD) with one or more cell penetrating peptides (CPPs). Methods are provided for genetically or otherwise modifying plants and for treating or preventing disease in plant cells comprising a cell wall.

Abstract: The present invention provides vectors that contain and express in vivo or in vitro sand fly Lu. longipalpis salivary antigens that elicit an immune response in animal or human against Leishmania, vaccine compositions comprising said vectors and/or Lu. longipalpis salivary polypeptides, methods of vaccination against Leishmania, and kits for use with such methods and compositions.

Abstract: The present invention relates to compositions and methods for stable transformation of green microalgae and for production of transgenic green microalgae and/or cyanobacteria that produce extremophile enzymes as co-products during the growth of the green microalgae and/or cyanobacteria for lipid biofuel production. Thus, the present invention provides nucleic acid constructs and methods of transformation useful in the production of stably transformed green microalgae and/or cyanobacteria expressing extremophile enzymes in combination with lipid production for biofuel.

Abstract: This present invention provides a method for monitoring ARV resistance, to determine viral fitness, and to forecast possible drug failure. The method provides improved personalized HIV/AIDS care to the patient-physician over existing assays at a reduced cost. This set of assays will utilize the same PCR amplicon of the patient HIV genome, which encompasses all of the drug targeted HIV-1 genes (polPR-RT-IN-envgp120-gp41) and not just PR-RT as with the prior systems. The greatest advantage of this method over previous is the rapid cloning of this amplicon into an HIV-1 genome vector through yeast recombination/gap repair. The vectors can be directly passed from yeast to mammalian cell line which has been specifically engineered to produce replication competent HIV-1 particles and to test susceptibility to all ARVs, i.e. PRIs, NRTIs, NNRTIs, T20, as well as entry and integrase inhibitors in development/clinical trials.

Abstract: The invention relates to a strain of the yeast Saccharomyces cerevisiae which, owing to deletion of the genomic sequences, no longer synthesizes hexose transporters and, as a consequence, can no longer grow on substrates with hexoses as the only carbon source, and whose ability of growing on a substrate with a hexose as the only carbon source is restored when it expresses a GLUT4 gene.

Abstract: The present invention relates to a nucleic acid comprising a Lactococcus CRISPR repeat region and/or a Lactococcus CRISPR spacer region.

Abstract: The present invention is related to a method for the selection of recombinant clones having integrated a gene of interest and a nucleotide sequence encoding a functional antidote protein to a toxic molecule, wherein said recombinant clones are the ones which survive following their integration into a host cell comprising in its genome a nucleotide sequence encoding said toxic molecule. The present invention is also related to a nucleic acid construct, a vector comprising said nucleic acid construct, a host cell and a cloning and/or sequencing kit for performing said method.

Abstract: Methods and compositions for targeted delivery of biotherapeutics are provided. The compositions comprise bile-sensitive St. thermophilus bacteria modified to release a biotherapeutic agent following bile exposure. Biotherapeutic agents released by the St. thermophilus bacteria disclosed herein include AQ and AQR rich peptides. Methods of the invention comprise administering to a subject a St. thermophilus bacterium modified to release a biotherapeutic agent following bile exposure. Administration of the St. thermophilus bacterium promotes a desired therapeutic response. The bacterium may be modified to express and release AQ or AQR rich peptides which subsequently inhibit cellular apoptosis or reduce mucosal damage. Thus, methods of the invention find use in treating or preventing a variety of gastrointestinal disorders including C. difficile infection and antibiotic-associated diarrhea.

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: A method for modulating cell differentiation capabilities using heterologous gene expression. Some embodiments of the invention relate to a method for inducing a cardiac progenitor cell by delivering a reprogramming factor to the cell, wherein the reprogramming factor comprises ETS2 or a combination of ETS2 and Mesp1.

Abstract: An isolated protein which is at least partially encoded by a polynucleotide sequence encoding a novel sulfotransferase is provided together with a composition which includes the isolated protein. A transgenic organism transformed by a polynucleotide encoding a protein which is at least partially encoded by a novel sulfotransferase is also provided. The invention also includes a process for in-vivo and in-vitro making a sulfated polysaccharide from an unsulfated polysaccharide or increasing the sulfur content of an already sulfated polysaccharide.

Abstract: The present invention includes a method to produce a recombinant mite Group 1 protein in a methyltrophic yeast or an Escherichia coli microorganism. The present invention also relates to a recombinant mite Group 1 protein obtained by such a method, such a recombinant protein being able to selectively bind IgE or cause proliferation of a T cell that proliferate in response to a native mite Group 1 protein. Also included in the present invention is the use of such a recombinant mite Group 1 protein to detect mite allergy or to reduce an allergic response to a mite Group 1 protein. The present invention also includes novel mite Group 1 nucleic acid molecules, proteins, recombinant molecules, and recombinant cells, as well as uses thereof.

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: Some aspects of this invention provide engineered microbes for oil production. Methods for microbe engineering and for use of engineered microbes are also provided herein. In some embodiments, microbes are provided that are engineered to modulate a combination of rate-controlling steps of lipid synthesis, for example, a combination of a step generating metabolites, acetyl-CoA, ATP or NADPH for lipid synthesis (a push step), and a step sequestering a product or an intermediate of a lipid synthesis pathway that mediates feedback inhibition of lipid synthesis (a pull step). Such push-and-pull engineered microbes exhibit greatly enhanced conversion yields and TAG synthesis and storage properties.

Abstract: The present invention relates to human Her-2/neu expressing plasmid constructs having anti-cancer activity and a DNA vaccine comprising same for preventing and/or treating cancer. The Her-2/neu DNA vaccines of the present invention can be effectively used as a therapeutic vaccine in reducing metastasis after tumor surgery or as a prophylactic vaccine for people with genetic high risk.

Abstract: Expression system of peptides on the bacterial surface characterized in that membrane-binding region the conserved sequence of the MSP1a protein of Anaplasma marginale.

Abstract: A synthetic gene devoid of CpG nucleotide derived by genetic engineering from copepod luciferases genes that code for a new secreted luciferase with a strong bioluminescent signal. This gene display advantageous properties to be used as a reporter genes in cell based assays.

Abstract: Disclosed are recombinant microorganisms for producing organic acids. The recombinant microorganisms express a polypeptide that has the enzymatic activity of an enzyme that is utilized in the pentose phosphate cycle. The recombinant microorganism may include recombinant Actinobacillus succinogenes that has been transformed to express a Zwischenferment (Zwf) gene. The recombinant microorganisms may be useful in fermentation processes for producing organic acids such as succinic acid and lactic acid. Also disclosed are novel plasmids that are useful for transforming microorganisms to produce recombinant microorganisms that express enzymes such as Zwf.

Abstract: This invention is related to bacterial engineering and the heterologous expression of useful compounds. In particular, the invention relates to a heterologous host that has been engineered for expression of a gene which is capable of polyketide or non-ribosomal peptide synthesis. Methods of treating cancer are also disclosed.

Abstract: Zymomonas is unable to synthesize pantothenic acid and requires this essential vitamin in growth medium. Zymomonas strains transformed with an operon for expression of 2-dehydropantoate reductase and aspartate 1-decarboxylase were able to grow in medium lacking pantothenic acid. These strains may be used for ethanol production without pantothenic acid supplementation in seed culture and fermentation media.

Abstract: The present invention generally relates to the production of industrially relevant quantities of selenoprotein enzymes in eukaryotic cell cultures. More specifically, the present invention generally relates to the production of such enzymes wherein one or more catalytic cysteine or serine residues are mutagenically replaced by selenocysteine.