Abstract: The present invention relates to methods for constructing a filamentous fungal strain for production of multiple recombinant polypeptides having biological activity. The present invention also relates to methods for producing multiple recombinant polypeptides having biological activity in a filamentous fungal strain. The present invention also relates to filamentous fungal strains expressing multiple recombinant polypeptides having biological activity.

Abstract: Provided herein are methods for assembling DNA fragments employing at least three enzymatic activities: DNA polymerase, flap endonuclease, and DNA ligase. Certain aspects include methods for generating closed circular DNA products, e.g., plasmid vectors, by assembling various DNA fragments having complementary ends that hybridize to one another. The resulting circular products can be introduced into host cells and selected for desired properties. Kits for performing the method are also provided.

Abstract: A biological material is disclosed for exerting antagonism against vegetable pathogens, which contains Bacillus mojavensis R3B mutant strain deposited under # NCAIM (P) B 001389 according to the Budapest Treaty. The biological material according to the invention is an effective antagonist against the pathogens of vegetables, preferably tomato, pepper, lettuce and/or cabbage, in particular against the pathogens selected from the group of Xanthomonas vesicatoria, Pseudomonas syringae and Clavibacter michiganensis vegetable pathogen bacteria and Pythium debaryanum, Phytophthora infestans, Alternaria alternata and Fusarium oxysporum vegetable pathogen fungi.

Abstract: A modified transposon vector and a method for introducing a foreign gene into a cell are provided.

Abstract: Isolated mutant Mycobacterium tuberculosis bacteria comprising a deletion in the ESAT-6 gene cluster region 3 (esx-3 region) are provided, as well as compositions comprising such, methods of production thereof and methods of use thereof.

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: Many fungal secondary metabolites are of industrial interest, such as antibiotics, while others are undesirable compounds such as mycotoxins. Overexpression of mtfA enhances production of fungal compounds with applications in the medical field, and overexpression or impaired mtfA expression decreases the production of compounds that negatively affect health/agriculture/economy such as mycotoxins.

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

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: 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 methods for constructing a filamentous fungal strain for production of multiple recombinant polypeptides having biological activity. The present invention also relates to methods for producing multiple recombinant polypeptides having biological activity in a filamentous fungal strain. The present invention also relates to filamentous fungal strains expressing multiple recombinant polypeptides having biological activity.

Abstract: Methods and means are provided for the exact exchange in eukaryotic cells, such as plant cells, of a target DNA sequence for a DNA sequence of interest through homologous recombination, whereby the selectable or screenable marker used during the homologous recombination phase for temporal selection of the gene replacement events can subsequently be removed without leaving a foot-print employing a method for the removal of a selected DNA flanked by two nucleotide sequences in direct repeats.

Abstract: The present invention discloses a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a replacement sequence of interest comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing two sets of DNA molecules of which the first set comprises DNA molecules each comprising a first non-functional fragment of the replacement sequence of interest flanked at its 5?-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence and the second set comprises DNA molecules each comprising a second non-functional fragment of the DNA replacement sequence of interest overlapping with the first non-functional fragment and flanked at its 3?-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence, wherein the first and second non-functional fragments

Abstract: The present invention is directed to a modified poxvirus vector that allows for the generation of recombinant poxviruses by a single recombination event. A modified poxvirus vector comprising at least one reporter gene located between two flanking sequences for homologous recombination is disclosed. Furthermore, a host cell comprising said vector and a method for the generation of recombinant poxviruses using said vector are provided.

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: A method for selecting a transformed yeast cell having integrated a nucleic acid fragment of interest by homologous recombination, cassettes and kits for carrying out the method are described.

Abstract: Exemplary methods for increasing TAG production in an algal cell during imbalanced growth conditions are provided. Some methods comprise knocking out an AOX gene, wherein the AOX gene produces an amino acid sequence having substantial similarity to the amino acid sequence of SEQ. ID. NO. 2. In further methods, the algal cell may be of genus Nannochloropsis. The AOX gene may be replaced by a construct having a nucleotide sequence having substantial similarity to SEQ ID. NOS. 3 through 5 (inclusive), wherein each of the sequences are next to or in close proximity to one another in a linear fashion. In some methods, the AOX gene may be replaced via homologous recombination. As a result, lipid production by the selected recombinant algal cell may be increased over that produced by a wild-type algal cell.

Abstract: The present invention relates to a process for producing a human-type glycoprotein having reduced glycosylation by genetically manipulating an enzyme involved in glycosylation using a Hansenula polymorpha system. In detail, the present invention relates to a process for producing a human-type glycoprotein by identifying a dolichyl-phosphate-mannose dependent ?-1,3-mannosyltransferase gene from H. polymorpha, constructing a H. polymorpha mutant strain producing a glycoprotein exhibiting reduced glycosylation by disrupting the identified gene, and subjecting the mutant strain to various genetic manipulations for the synthesis of human-type glycan.

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 invention concerns the use of a mutagenic agent blocking DNA replication in the cell for inserting in vitro a nucleic acid of interest inside a predetermined nucleotide sequence present in a chromosome contained in a prokaryotic or eukaryotic cell, said nucleic acid of interest being, prior to its insertion, included in a DNA vector which replicates in said prokaryotic or eukaryotic host cell.

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 present invention provides a method for producing an yeast having an increased cellulose hydrolysis ability. The method includes the step of introducing increased integration copy numbers of both a gene for an enzyme capable of hydrolyzing crystalline cellulose and a gene for an enzyme capable of hydrolyzing noncrystalline cellulose into a noncellulolytic yeast to give a transformed yeast. The yeast having an increased cellulose hydrolysis ability can be suitably used for ethanol production from cellulose-based materials.

Abstract: The present application relates to genetically modified yeasts for the production of glycoproteins having optimized and homogeneous glycan structures. These yeasts comprise an inactivation of the Och 1 gene, the integration by homologous recombination, into an auxotrophic marker, of an expression cassette comprising a first promoter, and an open reading frame comprising the coding sequence for an ?-1,2-mannosidase I, and the integration of a cassette comprising a second promoter different from said first promoter and the coding sequence for an exogenous glycoprotein. These yeasts make it possible to produce EPO with an optimized and 98% homogeneous glycosylation.

Abstract: Lambda phages that can be used to introduce recombineering functions into host cells are disclosed. Also disclosed are plasmids that can be used to confer recombineering functions to a variety of strains of E. coli and to other bacteria, including Salmonella, Pseudomonas, Cyanobacteria, Spirochaetes. These plasmids and phages can be isolated in vitro and can be used to transform bacterial cells, such as gram negative bacteria.

Abstract: This invention is directed to a general method for the chronic treatment, potential cure, or prevention of various metabolic and related diseases in people, including diabetes, by modulating IRS2 activity in cells and tissues in the body. IRS1 and IRS2 are part of the insulin or insulin-like growth factor signaling pathway. By upregulating the levels or functional activity of IRS2, insulin is used more efficiently by the body to control nutrient levels. By upregulating IRS2 levels or functional activity in pancreatic ?-cells, glucose sensing and insulin secretion are enhanced.

Abstract: The present invention relates to the field of the veterinary medicine of bovine animals. In particular the invention relates to a recombinant Trypanosoma theileri parasite, preferably comprising a heterologous nucleic acid sequence that is capable of encoding a protein for instance an antigen, a cytokine, a hormone, an antimicrobial protein, or an antibody. Also disclosed are uses of and methods for making and using the recombinant T. theileri parasite in medical or non-curative treatments; in particular as a sustained delivery vector for proteins to bovine animals, e.g. as a vaccine.

Abstract: Methods for enhancing single cross-over homologous recombination in gram positive bacteria are presented. These methods provide enhanced capability to genetically modify gram positive bacteria.

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

Abstract: The invention relates to a set of genetic constructs which allow the efficient and reproducible introduction of a specific nucleotide sequence at a fixed position in the genome by generating a double strand break at a specific position in the genome using a meganuclease and so stimulating a homologous recombination event at this locus between the genomic site and a transfected donor sequence. The present invention also relates to methods using these constructs and to these materials in the form of a kit.

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: This disclosure describes genetically modified photosynthetic microorganisms, including Cyanobacteria, that contain one or more exogenous genes encoding a diacyglycerol acyltransferase, a phosphatidate phosphatase, and/or an acetyl-CoA carboxylase, and which are capable of producing increased amounts of fatty acids and/or synthesizing triglycerides.

Abstract: The present invention relates to an attenuated Salmonella typhi having mutation in chromosomal gene loci, its use as a potent vaccine candidate to combat the Salmonella infection.

Abstract: The present invention relates to nucleotide sequences, which are variants of att recombination sequences, involved in sequence-specific recombination of DNA in eukaryotic cells, whereby sequence specific recombination is performed by a bacteriophage lambda integrase Int. Such att recombination sequences being e.g. attP.b, attP.a, attL.a, attR.a and attR.b. The present invention further relates to a method of sequence-specific recombination of DNA in eukaryotic cells, comprising the introduction of a first DNA comprising a nucleotide sequence containing at least one recombination sequence into a cell, introducing a second DNA comprising a nucleotide sequence containing at least one further recombination sequence into a cell, and performing the sequence specific recombination by a bacteriophage lambda integrase Int, whereby at least one of said first or second DNAs is an att recombination sequence being e.g. attP.b, attP.a, attL.a, attR.a or attR.b.

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

Abstract: The present disclosure relates to methods of integrating recombinant polynucleotides into genomes of unicellular organisms. In particular, the present disclosure relates to the modified unicellular organisms that contain integrated recombinant polynucleotides in their genomes and methods for production of commodity chemicals by the use of such organisms.

Abstract: The present invention relates to a method for isolating from the immunological gene repertoire a gene coding for a receptor having the ability to bind a preselected ligand. Receptors produced by the gene isolated by the method, particularly catalytic receptors, are also contemplated.

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: Auxotrophic Agrobacterium and methods employing auxotrophic Agrobacterium are provided. Auxotrophic Agrobacterium may be used in a variety of methods including biologically containing Agrobacterium comprising a transgene and transforming a plant cell without using an Agrobacterium counter-selective agent. Transforming maize immature embryos using an Agrobacterium auxotrophic for thymidine results in comparable transformation efficiency as transformation achieved using prototrophic Agrobacterium. Methods for producing an Agrobacterium thymidine auxotroph and using the auxotroph in transformation methods are disclosed. Transformed tissues and plants produced using methods of the present invention are also provided.

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 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 provides a unidirectional site-specific integration system for integrating a nucleic acid into the genome of a target cell. The provided system includes a site-specific integrating expression cassette (INTEC) vector, consisting of (a) a polynucleotide of interest operably linked to a promoter, (b) a single recombination site, and (c) a hybrid recombination site. In using the subject systems for site-specific integration, the INTEC vector and integrase are introduced into the target cell and the cell is maintained under conditions sufficient to provide for site-specific integration of the nucleic acid into the target cell genome via a recombination event mediated by the site-specific recombinase. Also provided are kits that include the subject systems. The subjects systems, methods and kits find use in a variety of different applications, several representative ones of which are described in detail as well.

Abstract: The present invention provides an improved expression system for the production of recombinant polypeptides utilizing auxotrophic selectable markers. In addition, the present invention provides improved recombinant protein production in host cells through the improved regulation of expression.

Abstract: Described herein are baculovirus vectors, host cells containing the baculovirus vectors, methods of using the baculovirus vectors for cloning genes, and kits containing the baculovirus vectors.

Abstract: The present invention describes an L-glutamic acid-producing bacterium which belongs to the genus Pantoea, Enterobacter, Klebsiella or Erwinia, wherein the bacterium has been modified by gene recombination to inactivate the rpoS gene. A method is also described for culturing the bacterium in a medium to cause accumulation of L-glutamic acid in the medium, and collecting L-glutamic acid from the medium.

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: The present invention is related to a new method for replacing or deleting DNA sequences in Clostridia, with high efficiency, easy to perform and applicable at an industrial level. This method is useful to modify several genetic loci in Clostridia in a routine manner. This method is based on a replicative vector carrying at least two marker genes.

Abstract: A method for producing a chimaeric human papillomavirus (HPV) L1 polypeptide containing a heterologous peptide, and in particular, a HPV L2 peptide comprising the steps of introducing a DNA sequence coding for the heterologous peptide into a DNA sequence coding for the L1 polypeptide; introducing the DNA sequence including the sequences for the L1 polypeptide and heterologous peptide into a host cell in which the DNA sequence can be expressed; causing expression of the DNA sequence; and recovering the resulting chimaeric L1 polypeptide which includes the heterologous peptide. The invention also describes a vector for use in the method, a host cell containing the vector, and a vaccine including the chimaeric HPV L1 polypeptide produced according to the method.

Abstract: The present invention relates to methods for increasing homologous recombination of a nucleic acid sequence introduced into a host cell, comprising: (a) introducing into a population of filamentous fungal host cells a first nucleic acid sequence encoding a recombination protein and a second nucleic acid sequence comprising one or more regions which are homologous with the genome of the filamentous fungal host cell, wherein (i) the recombination protein promotes the recombination of the one or more regions with the corresponding homologous region in the host's genome to incorporate the second nucleic acid sequence by homologous recombination, and (ii) the number of host cells comprising the incorporated second nucleic acid sequence in the population is increased at least 20% compared to the same population without the first nucleic acid sequence; (b) and isolating from the population a filamentous fungal cell comprising the incorporated second nucleic acid sequence.

Abstract: T4 bacteriophages are bound to substrates such as liposomes using a binder.

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.