Abstract: The present invention relates to novel engineered lower eukaryotic host cells for expressing heterologous proteins and to methods of generating such strains.

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: A host-vector system that uses the RNA-based copy number control mechanism of ColE1-type plasmids for regulating the expression of a marker gene allows for antibiotic-free selection of plasmids and is useful for production of plasmid DNA and recombinant proteins.

Abstract: The present invention is directed to the detection of problematic foaming and bulking bacterial species in the biological wastewater treatment process. The invention provides various compositions of matter and methods for the detection of foaming and bulking bacterial species and genera in wastewater and other samples. PCR primers capable of amplifying 16s rRNA gene sequences from various foaming and bulking bacterial species are provided, as are probes that will specifically hybridize with PCR amplification products produced by the disclosed primers. In certain embodiments, the use of the disclosed PCR primers and probes in detection assays is disclosed.

Abstract: Methods are provided for determining whether a subject has a graft tolerant phenotype. In practicing the subject methods, the expression level of one or more gene in a sample from the subject, e.g., a blood sample, is assayed to obtain a gene expression result, where the gene expression result includes a result for a biomarker of graft tolerance. The obtained gene expression result is then employed to determine whether the subject has a graft tolerant phenotype. Also provided are compositions, systems and kits that find use in practicing the subject methods. The methods and compositions find use in a variety of applications, including the determination of an immunosuppressive therapy regimen.

Abstract: Vectors expressible in a host that is the rhaBAD promoter region of the L-rhamnose operon operably linked to a transcriptional unit that is: a) a nucleic acid sequence which is heterologous to the host, and b) a prokaryotic signal sequence operably linked to the nucleic acid sequence. The prokaryotic signal sequence is selected from signal peptides of periplasmatic binding proteins for sugars, amino acids, vitamins and ions. The expression of the nucleic acid sequence is controlled by the promoter region. The vector is used for the regulated heterologous expression of a nucleic acid sequence in a prokaryotic host. This is an isolated and purified nucleic acid sequence expressible in a host is the promoter region of the L-rhamnose operon. There is a method for producing a polypeptide in a host using the vector.

Abstract: Methods for the production of unicellular photosynthetic organisms capable of producing cell adhesion proteins are disclosed. DNA constructs as well as methods for integration of the DNA constructs into the genomes of unicellular photosynthetic organisms for the expression of cell adhesion proteins are also disclosed.

Abstract: The invention provides methods of preparing bacteria of interest with a stably integrated nucleic acid sequence of interest.

Abstract: The present invention relates to methods and compositions for the expression of protein. Embodiments of methods may comprise the cleavage and repair of a nucleotide sequence encoding a highly expressed protein leading to a reduction in the expression of the highly expressed protein.

Abstract: The present invention is based, at least in part, on the development of a mating-based yeast two-hybrid screen that allows simultaneous screening for mutations that disrupt yeast two-hybrid interactions between a protein and multiple interacting partners. By coupling PCR mutagenesis and homologous recombination/gapped plasmid repair with a mating-based assay, the present invention allows screening for unique mutations that disrupt interaction with one partner, but not others. It also allows identification of specific mutations that may lie at protein-protein interfaces common to two or more partners, without employing multiple rounds of screening. In addition to screening against multiple interacting partners, the present invention removes the need for a two-step selection because truncations, frameshifts, or any mutations that affect folding are eliminated as disruptions that affect all protein partners.

Abstract: The present invention relates to a method for producing a protein of interest containing one or more disulfide bonds in its native state. The method comprises that a prokaryotic host cell is genetically engineered to express the protein of interest and a sulfhydryl oxidase in the cytoplasm of the host cell. The protein of interest is formed in a soluble form and contains disulfide bonds due to the presence of the sulfhydryl oxidase in the cytoplasm of said host cell. The present invention relates also to a prokaryotic host cell and a vector system for producing a protein of interest containing natively folded disulfide bonds.

Abstract: A system for classifying thyroid nodule tissue as malignant or benign is provided that is based on the identification of sets of gene transcripts, which are characterized in that changes in expression of each gene transcript within a set of gene transcripts can be correlated to with either malignant or benign thyroid nodule disease. The thyroid classification system provides for sets of “thyroid classifying” target sequences and further provides for combinations of polynucleotide probes and primers derived there from. These combinations of polynucleotide probes can be provided in solution or as an array. The combination of probes and the arrays can be used for diagnosis. The invention further provides further methods of classifying thyroid nodule tissue.

Abstract: This invention relates to chromosomal engineering via DNA repair process. The process of the invention comprises the steps of: 1) submitting at least one source of biological activity, e.g. Deinococcus radiodurans, to radiation, desiccation and/or chemical treatment liable to damage the DNA, so as to substantially shatter its chromosomes into short fragments; 2) annealing complementary single strand tails extended by the synthesis templated on partially overlapping DNA fragments of said shattered chromosomes; 4) converting the resulting long linear DNA intermediates into intact circular chromosomes, by means of a RecA dependent homologous recombination; whereas at least one foreign source of genetic material, e.g. DNA, can be introduced during steps 2 and/or 3; and 4) optionally separating and collecting the recombined chromosomes thus obtained.

Abstract: The invention relates to modifying plasmid origins of replication to create hybrid origins of replication containing nucleotide sequences from more than one plasmid. The invention also relates to a modified origin of replication cassette that is portable or exchangeable due to the creation of multiple cloning sites flanking the origin of replication. Methods and plasmids for use in exchanging origins of replication are disclosed. Such modified or hybrid plasmids provide useful cloning tools that allow for regulation of the level of expression of a desired protein.

Abstract: The present invention relates to systems which harness the molecular biology of a living cell to direct evolution of biological entities of interest. According to the invention, a host cell is engineered to facilitate mutation of a nucleic acid target corresponding to that entity and select for desirable mutants. As applied to populations of host cells, the invention provides a means to generate and contemporaneously select mutants of interest, allowing for the production of extremely diverse libraries enriched in the most ‘fit’ mutants.

Abstract: The present invention relates to a vector expressible in a prokaryotic host and a nucleic acid sequence comprising a mannose-inducible promoter of the mannose operon of Bacillus subfiles wherein the vector and nucleic acid sequence, respectively, can be suitably used for transforming a host cell for expression of a heterologous nucleic acid sequence coding a polypeptide in, in particular, high cell density fermentation.

Abstract: The present invention relates to animal protein free cell culture media comprising a combination of non-animal derived peptides derived from soy hydrolysate and yeast hydrolysate. The invention also provides an animal protein free culture process, wherein cells are cultivated, propagated and passaged without animal-derived components. This process is useful for cultivating cells, such as recombinant cells or cells infected with a virus, and for production biological products by cell culture processes under conditions devoid of animal protein components.

Abstract: Disclosed are tyrosine-modified rAAV vectors, as well as infectious virions, compositions, and pharmaceutical formulations that comprise them. Also disclosed are methods of preparing and methods for using the disclosed tyrosine-phosphorylated capsid protein mutant rAAV vectors in a variety of diagnostic and therapeutic applications including in vivo and ex vivo gene therapy, and large-scale production of rAAV vectors.

Abstract: Aspects of the invention include inducible expression systems in which a transcription modulator having a distributed protein transduction domain is employed. Aspects of the invention further include methods of using the systems to induce expression of a coding sequence, as well as kits that find use in practicing methods of the invention. The systems, components thereof, methods and kits find use in a variety of different applications.

Abstract: Methods are described that bias cells, such as potent and multipotent stem cells, by transfection with a nucleic acid sequence, to differentiate to a desired end-stage cell or a cell having characteristics of a desired end-stage cell. In particular embodiments, human neural stem cells are transfected with vectors comprising genes in the homeobox family of transcription factor developmental control genes, and this results in a greater percentage of resultant transformed cells, or their progeny, differentiating into a desired end-stage cell or a cell having characteristics of a desired end-stage cell.