Abstract: Described herein are methods for increasing the amount of protein secreted by a cell. In one case, a cell is provided which contains a heterologous nucleic acid encoding a protein having unfolded protein response modulating activity and a heterologous nucleic acid encoding a protein of interest to be secreted. In one case, the protein having unfolded protein response modulating activity is selected from the proteins selected from the group consisting of HAC1, PTC2 and IRE1. The protein of interest can be any secreted protein such as a therapeutic or an industrial enzyme. For example the protein can be selected from the group consisting of lipase, cellulase, endo-glucosidase H, protease, carbohydrase, reductase, oxidase, isomerase, transferase, kinase, phosphatase, alpha-amylase, glucoamylase, lignocellulose hemicellulase, pectinase and ligninase.

Abstract: The present invention describes the genetic engineering of production microorganisms used in biotechnology to improve their properties so that they produce industrially useful products more efficiently from fermentable sugars derived from biomass. The engineered microorganisms endowed with functional coupling of oxidation and reduction of substrates by dehydrogenases requiring pyridine nucleotides (NAD/NADH) result in simultaneous enhancement of reduction potential enzyme activity involving the transfer of electrons. In particular, this invention relates to the construction of an excisable gene expression cassette for expression of two different dehydrogenases leading to enhanced production of ethanol.

Abstract: New vectors expressible in a host comprising the promoter region of the melibiose operon operably linked to a transcriptional unit that includes nucleic acid sequence which is heterologous to the host. The expression of the nucleic acid sequence is controlled by the promoter region of the melibiose operon. The new vector can be used for the regulated heterologous expression of a nucleic acid sequence in a prokaryotic host. There is an isolated and purified nucleic acid sequence expressible in a host comprising the promoter region of the melibiose operon operably linked to a transcriptional unit that includes a nucleic acid sequence which is heterologous to the host. The expression of the nucleic acid sequence is controlled by the promoter region of the melibiose operon. A prokaryotic host is transformed with the vector or the isolated and purified nucleic acid sequence. There is a method for producing a polypeptide in a host using the vector.

Abstract: The present invention relates to the use of nucleic acid sequences for regulating the transcription and expression of genes, the novel promoters and expression units themselves, methods for altering or causing the transcription rate and/or expression rate of genes, expression cassettes comprising the expression units, genetically modified microorganisms with altered or caused transcription rate and/or expression rate, and methods for preparing biosynthetic products by cultivating the genetically modified microorganisms.

Abstract: The invention encompasses methods of using quantitative PCR to detect fungal organisms in clinical and environmental samples and to generate standards that allow the quantification of fungal organisms in the samples.

Abstract: A system and method for heterologous expression of polyketide biosynthetic pathways from streptomycetes hosts in Escherichia coli for the production and discovery of secondary metabolites. Genomic DNA from Streptomyces rimosus encoding the oxytetracycline biosynthetic pathway is inserted into the genome of the surrogate host Myxococcus xanthus. The M. xanthus transcriptional machinery recognizes and uses the streptomycetes promoter regions to express the biosynthetic enzymes. Co-expression in E. coli of S. rimosus oxytetracycline biosynthesis enzymes and M. xanthus ?54, a key piece of the M. xanthus transcriptional machinery, enables E. coli to recognize and use the promoters from the S. rimosus oxytetracycline biosynthetic pathway, facilitating production of oxytetracycline.

Abstract: Methods are provided for the synthesis and secretion of recombinant hetero-multimeric proteins in mating competent yeast. A first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

Abstract: The invention provides a target and methods for specific binding and inhibition of RNAP from bacterial species. The invention is directed to a method for identifying agents that bind to a bacterial RNAP homologous RNA-exit-channel amino-acid sequence, comprising preparing a reaction solution comprising the agent to be tested and an entity comprising a bacterial RNAP homologous RNA-exit-channel amino-acid sequence, and detecting presence or amount of binding. The invention has applications in control of bacterial gene expression, control of bacterial growth, antibacterial chemistry, and antibacterial therapy.

Abstract: The present invention relates to methods of obtaining genetic competence in non-competent Bacillus cells for their transformation with exogenous DNA.

Abstract: Methods are provided for producing highly purified compositions of nucleic acids by using a highly streamlined and readily automated process. The methods use static mixers for lysing cells and precipitating debris, following by centrifugation and ion exchange chromatography. The process may include a purification step using tangential flow ultrafiltration. A scaleable process for producing pharmaceutical grade plasmid DNA, useful for gene therapy, is provided.

Abstract: Disclosed herein are ammonia-specific 5?-XMP aminase mutants and a method for preparing the same. A mutation is introduced into the active site of glutamine-dependent catalysis in 5?-XMP aminase. The resulting 5?-XMP aminase mutant is devoid of the glutamine-dependent activity and specifically reacts with external ammonia in converting 5?-XMP into 5?-GMP. Thus, the ammonia-specific 5?-XMP aminase mutant is stabler within cells compared to the wild type, and can be useful in the industrial conversion of 5?-XMP into 5?-GMP.

Abstract: Inhibitors of the tmRNA pathway have antibacterial activity with broad species specificity, including B. anthracis and other pathogens of military and civilian interest. Identified cyclic or linear peptides are further selected by in vivo selection methods, kill bacterial pathogens when added exogenously, and/or eliminate plasmids carrying antibiotic resistance or virulence genes. The molecular target of each cyclic peptide is in the tmRNA pathway and the tmRNA pathway is inhibited in vitro and in vivo by the addition of the peptides.

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: The present invention provides a modified promoter DNA capable of enhancing transcription of genes encoding proteins or polypeptides, and a method for producing proteins or polypeptides efficiently by use of the modified promoter DNA. A promoter DNA recognized by SigA and SigE, which is produced by modifying a nucleotide sequence including a promoter recognized by SigA and bases in the vicinity thereof; an expression vector harboring the promoter DNA; a recombinant microorganism containing the expression vector; and a method for producing proteins or polypeptides characterized by culturing the recombinant microorganism.

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: Methods are disclosed for producing proteins having biological activity for blood coagulation mediated by Factor VIIa or Factor IX. The proteins are produced by mammalian host cells which have been stably transfected with a DNA construct containing a nucleotide sequence which codes at least partially for either Factor VII or Factor IX. The nucleotide sequence comprises a first nucleotide sequence encoding a calcium binding domain, joined to a second nucleotide sequence positioned downstream of the first sequence. The second sequence encodes a catalytic domain for the serine protease activity of either Factor VIIa or Factor IX. The joined sequences code for proteins having substantially the same biological activity for blood coagulation as either Factor VIIa or Factor IX.

Abstract: Devices are disclosed for serologically detecting an infection with human-pathogenic Yersinia ssp, wherein said device comprises at least one antigen selected from the group of antigens consisting of the following group: YopD, YopH, YopM, YopE, V-AG and YopN or a fragment of one of said antigens having at least eight consecutive amino acids and furthermore one of two proteins selected from MyfA and PsaA or fragments of one of said two proteins having at least eight consecutive amino acids.

Abstract: The present invention relates to nucleic acids and nucleic acid fragments encoding amino acid sequences for flavonoid biosynthetic enzymes in plants, and the use thereof for the modification of flavonoid biosynthesis in plants. More particularly, the flavonoid biosynthetic enzyme is selected from the group consisting of chalcone isomerase (CHI), chalcone synthase (CHS), chalcone reductase (CHR), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductase (LCR), flavonoid 3?,5? hydrolase (F3?5?H), flavanone 3-hydroxylase (F3H), flavonoid 3?-hydroxylase (F3?H), phenylalanine ammonia-olyase (PAL) and vestitone reductase (VR), and functionally active fragments and variants thereof.

Abstract: This invention provides a modified vaccinia topoisomerase enzyme containing an affinity tag which is capable of facilitating purification of protein-DNA complexes away from unbound DNA. This invention further provides a modified sequence specific topoisomerase enzyme. This invention provides a method of ligating duplex DNAs, a method of molecular cloning of DNA, a method of synthesizing polynucleotides, and a method of gene targeting. Lastly, this invention provides a recombinant DNA molecule composed of segments of DNA which have been joined ex vivo by the use of a sequence specific topoisomerase and which has the capacity to transform a suitable host cell comprising a DNA sequence encoding polypeptide activity.

Abstract: Methods and compositions for detecting molecular interactions are provided. Aspects of the invention include the use of a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.