Abstract: The present invention relates to a targeting system comprising, preferably as distinct components (a) a transposon which is devoid of polynucleotide encoding a functional transposase comprising (aa) a polynucleotide of interest and; (ab) a DNA sequence specifically recognized by a DNA binding domain; and (ba) a fusion protein comprising (i) said DNA binding domain; or (ii) a (poly)peptide binding domain binding to a (poly)peptide comprising said DNA binding domain; and (iii) a DNA targeting domain; or (iv) a (poly)peptide binding domain that binds to a cellular or engineered (poly)peptide that comprises a DNA targeting domain; or (bb) a polynucleotide encoding the fusion protein of (ba); and (ca) a transposase or a fragment or derivative thereof having transposase function; or (cb) a polynucleotide encoding the transposase or fragment or derivative thereof having transposase function of (ca).

Abstract: The present invention concerns a method for improving growth characteristics of plants by increasing expression and/or activity in a plant of an LRR receptor kinase or a homologue thereof. One such method comprises introducing into a plant an RLK827 nucleic acid molecule or functional variant thereof. The invention also relates to transgenic plants having improved growth characteristics, which plants have modulated expression of a nucleic acid encoding an LRR receptor kinase. The present invention also concerns constructs useful in the methods of the invention.

Abstract: This invention relates to an isolated protein MONaKA and methods of its use. Specifically, the invention is directed to a protein that modulates the Na,K-ATPase and glutamate transporters, by binding to the b-subunit of the plasma membrane Na,K-ATPase (Na pump).

Abstract: The present invention provides a method for constructing recombinant translationally coupled operons, a method for producing useful metabolites using the bacterium containing the coupled operons, and a method for monitoring gene expression.

Abstract: The present invention provides duplexed parvovirus vector genomes that are capable under appropriate conditions of forming a double-stranded molecule by intrastrand base-pairing. Also provided are duplexed parvovirus particles comprising the vector genome. Further disclosed are templates and methods for producing the duplexed vector genomes and duplexed parvovirus particles of the invention. Methods of administering these reagents to a cell or subject are also described. Preferably, the parvovirus capsid is an AAV capsid. It is further preferred that the vector genome comprises AAV terminal repeat sequences.

Abstract: The present invention relates generally to the fields of molecular biology and protein technology. More specifically, the invention concerns signal sequences for the secretion of heterologous polypeptide from bacteria. The invention also concerns recombinant polypeptides and uses thereof.

Abstract: The invention relates to a nucleic acid preparation with a content of below 1% protein, preferably below 0.1% protein, free of ethidium bromide, phenol, cesium chloride and detergents based on octyl phenol poly(ethylene glycol ether)n and with a content of below 1 EU/mg DNA of endotoxins. Said preparation is suitable as a drug particularly in gene therapy.

Abstract: This invention provides a new approach to the design of a virus with a defective replication cycle, which can be rescued by wild type virus co-infection, and which expresses foreign antigenic epitopes that contribute to the elimination of virus infected cells and then to viral clearance. The vector of the invention, by expression of epitopes derived from common pathogens, by-passes existing tolerance of virus specific T cell responses. The vector will only replicate in virus infected cells.

Abstract: A gene construct comprising a programmed-cell-death executioner gene having a nuclear localization signal, a deleted signal peptide, an inhibitor-resistant binding site, a promoter, and an activator. A method of making a gene construct, by modifying a programmed-cell-death executioner gene by adding a nuclear localization signal, deleting a signal peptide, mutating a binding site for an inhibitor to make it inhibitor-resistant, adding a promoter for exclusive expression in selected cells, and adding an activator. A method of eliminating undesired cells from a patient. A method of treating cancer. An array comprising at least two gene constructs wherein all of the gene constructs differ with respect to the programmed-cell-death executioner gene and the nuclear localization signal. A method of personalizing anti-cancer treatment. A method of increasing DNase 1 resistance to actin binding. A method of increasing catalytic activity of DNase 1 binding.

Abstract: The present invention provides improved chimeric glycoproteins (GPs) and improved lentiviral vectors pseudotyped with those glycoproteins. Also provided are methods and compositions for making such glycoproteins and vectors, and improved methods of in vitro and in vivo transduction of cells with such vectors. Improved chimeric GPs encode the extracellular and transmembrane domains of GALV or RD114 GPs fused to the cytoplasmic tail of MLV-A GP. Vectors pseudotyped with these GAL V/TR and RD 114/TR GP chimeras have significantly higher titers than vectors coated with the parental GPs. Additionally, RD114/TR-pseudotyped vectors are efficiently concentrated and are resistant to inactivation induced by the complement of both human and macaque sera. RD114 GP-pseudotyped lentiviral vectors have particular utility for in vivo gene transfer applications.

Abstract: Polypeptides, polynucleotides, methods, compositions, and vaccines comprising influenza hemagglutinin and neuraminidase variants are provided.

Abstract: The present invention provides a method and compositions for high throughput screening of genetically modified photosynthetic organisms for plasmic state. The present invention provides methods of producing one or more proteins, including biomass degrading enzymes in a plant. Also provided are the methods of producing biomass degradation pathways in alga cells, particularly in the chloroplast. Single enzymes or multiple enzymes may be produced by the methods disclosed. The methods disclosed herein allow for the production of biofuel, including ethanol.

Abstract: A seed-specific expression vector and its construction methods and applications are disclosed. A fusion protein expression cassette consisting of Arachis hypogaea oleosin gene-apolipopoprotein A-IMilano (A-IM) gene driven by Brassica napus oleosin gene promoter is inserted between the HindIII and SacI sites of a plant binary expression vector pBI121, obtaining the plant expression vector pBINOA of the invention. In addition, a method for producing apolipoprotein A-IMilano is provided, in which the expression vector is used to transform oil sunflower which is used as a plant bioreactor. The method can not only improve the yield of apolipoprotein A-IMilano, but also greatly reduce production costs, and is suitable for industrial production.

Abstract: The present invention relates to the fields of biotechnology and molecular biology. In particular, the present invention relates to the construction and use of nucleic acid molecules comprising cloning sites which differ in nucleotide sequence. In particular embodiments, the present invention relates to nucleic acid molecules which contain recombination sites with different primer binding sites. These different primer binding sites may be used to sequence different ends of nucleic acid segments located between the two recombination sites.

Abstract: Methods are provided for adding a terminal sequence tag to nucleic acid molecules for use in RNA or DNA amplification. The tag introduced may be used as a primer binding site for subsequent amplification of the DNA molecule and/or sequencing of the DNA molecule and therefore provides means for identification and cloning of the 5?-end or the complete sequence of mRNAs.

Abstract: A method of producing recombinant plasmid DNA using substantially solid growth medium and disposable vessels in place of conventional liquid fermentation processes. The method includes inoculating a host organism containing the recombinant plasmid DNA onto the substantially solid growth medium in a disposable vessel; allowing the host organism to grow on the growth medium under conditions conducive to such growth; removing the host organism from the growth medium and lysing the host organism to access the recombinant plasmid DNA; and purifying the recombinant plasmid DNA.

Abstract: A method of creating a biotechnological product and an efficient and stable bio-luminescence vector which could be used for tracking Gram-negative bacteria when distributing inside animal body are provided. Through conjugation, this auto-luminescence vector can be easily transmitted from bacteria to bacteria among Gram-negative bacteria, and may facilitate bacteria to be luminescence-labeled for subsequently analyzing the dynamic change of bio-luminescent bacteria within animal body in vivo. This system includes a lacZ promoter-driven luxABCDE, a high copy number of ColE1 replicon, and a high plasmid stability of the conjugative and broad host-ranged plasmid pSE34 from Salmonella enterica serovar Enteritidis Sal550. This resulting construct pSE-Lux1 can not only conjugatively transmit among bacteria with broad host range, but also stably maintain in bacteria to efficiently express the bio-luminescent luxABCDE without supplementing the subtract for luciferases and the antibiotics for plasmid selection.

Abstract: Adipose cells (sebocytes) are described, The invention especially relates to sebaceous gland cells and to a sebaceous gland cell line with the property of being continuously grown over many sub-cultures. The sebocytes are excellently suited for useful applications.

Abstract: The present invention provides a polypeptide comprising (i) a leader sequence, the leader sequence comprising a (a) secretion pre sequence, and (b) the following motif: -X1-X2-X3-X4-X5- where X1 is phenylalanine, tryptophan, or tyrosine, X2 is isoleucine, leucine, valine, alanine or methionine, X3 is leucine, valine, alanine or methionine, X4 is serine or threonine and X5 is isoleucine, valine, alanine or methionine; and (ii) a desired protein heterologous to the leader sequence. A polypeptide of the invention may additionally comprise, as part of the leader sequence, a secretion pro sequence. The invention also provides a polynucleotide comprising a sequence that encodes a polypeptide of the invention and a cell, preferably a yeast cell, comprising said polynucleotide.

Abstract: A method of secretory expression of lysostaphin in Escherichia coli at high level, which comprises constructing a expression vector by cloning a sequence encoding a signal peptide which is suitable for secretory expression in Escherichia coli before part or whole gene sequence which encodes mature lysostaphin, and ligating the cloned sequence with a promoter; and transforming Escherichia coli with the expression vector, culturing and fermenting, and then isolating lysostaphin from the supernatant of the fermentation broth. The advantage of secretory expression is that the expression product can exist in the medium in an active form, and thus does not need the process for renaturation of the inclusion body; it is more easily to purify from the supernatant of the fermentation broth with high rate of recovery; and there is less contamination from the host's proteins.

Abstract: The present invention relates to novel expression cassettes and vectors for efficiently producing authentic recombinant human proteins from stable cultures of novel human cell lines, the authentic recombinant proteins produced therefrom, and antibodies raised against those authentic recombinant proteins.

Abstract: The present invention relates to DNA loaded gold nanoparticles embedded in sharp carbonaceous carriers useful for higher DNA delivery efficiently into plants. These nanogold embedded carbon matrices are prepared by heat treatment of biogenic intracellular gold nanoparticles. The DNA delivery efficiency is tested on model plants. These materials reveal good dispersion of the transport material, producing a greater number of GUS foci per unit area. The added advantages of the composite carrier are the lower plasmid and gold requirements. Plant cell damage with the prepared carbon supported particles is very minimal and can be gauged from the increased plant regeneration and transformation efficiency compared to that of the commercial micrometer sized gold particles. This can be attributed to the sharp edges that the carbon supports possess, which lead to better piercing capabilities with minimum damage.

Abstract: This invention provides a series of low-copy number plasmids comprising restriction endonuclease recognition sites useful for cloning at least three different genes or operons, each flanked by a terminator sequence, the plasmids containing variants of glucose isomerase promoters for varying levels of protein expression. The materials and methods are useful for genetic engineering in microorganisms, especially where multiple genetic insertions are sought.

Abstract: The present invention provides recombinant bicistronic flaviviruses, particularly live attenuated recombinant bicistronic flavivirus, which comprise, in order from 5? to 3?, a viral 5?UTR, an ORF encoding all viral proteins, an internal ribosome entry site, an exogenous nucleotide sequence that encodes an exogenous polypeptide, and a viral 3?UTR. Infection of a host cell with a recombinant flavivirus provides for expression of the exogenous nucleic acid in a host cell. Such recombinant flavivirus are useful for delivering a protein to a mammalian host; and for eliciting an immune response to the exogenous polypeptide.

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: Methods and compositions for making stable recombinant yeast 2 ?m plasmids are provided. Homologous recombination is performed to clone a nucleic acid of interest into the yeast 2 ?m plasmid. Heterologous nucleic acid subsequences are recombined between an FLP and a REP2 gene of the plasmid.

Abstract: The invention relates to kinase ligands and polyligands. In particular, the invention relates to ligands, homopolyligands, and heteropolyligands that modulate protein kinase D (PKD) activity. The ligands and polyligands are utilized as research tools or as therapeutics. The invention includes linkage of the ligands and polyligands to cellular localization signals, epitope tags and/or reporters. The invention also includes polynucleotides encoding the ligands and polyligands.

Abstract: A method of creating a biotechnological product and an efficient and stable bio-luminescence vector which could be used for tracking Gram-negative bacteria when distributing inside animal body are provided. Through conjugation, this auto-luminescence vector can be easily transmitted from bacteria to bacteria among Gram-negative bacteria, and may facilitate bacteria to be luminescence-labeled for subsequently analyzing the dynamic change of bio-luminescent bacteria within animal body in vivo. This system includes a lacZ promoter-driven luxABCDE, a high copy number of ColE1 replicon, and a high plasmid stability of the conjugative and broad host-ranged plasmid pSE34 from Salmonella enterica serovar Enteritidis Sal550. This resulting construct pSE-Lux1 can not only conjugatively transmit among bacteria with broad host range, but also stably maintain in bacteria to efficiently express the bio-luminescent luxABCDE without supplementing the subtract for luciferases and the antibiotics for plasmid selection.

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 invention provides a method and compositions for high throughput screening of genetically modified photosynthetic organisms for plasmic state. The present invention provides methods of producing one or more proteins, including biomass degrading enzymes in a plant. Also provided are the methods of producing biomass degradation pathways in alga cells, particularly in the chloroplast. Single enzymes or multiple enzymes may be produced by the methods disclosed. The methods disclosed herein allow for the production of biofuel, including ethanol.

Abstract: It is intended to provide a drug to be used in gene therapy which specifically targets abnormal cells such as tumor cells and destroys the same for healing. Namely, a drug comprising, as the active ingredient, a proliferative vector which contains a Survivin promoter proliferating depending on the expression of Survivin. This drug may be used in order to treat tumor. In this drug, use may be made of an adenovirus as the vector. In the adenovirus of this drug, an endogenous promoter of an E1A domain may be substituted with a Survivin promoter.

Abstract: The present invention relates to a purified nucleic acid sequence encoding a homologue of human interleukin 10 (IL-10), wherein said IL-10 homologue is expressed during the latent phase of infection by a virus of the herpesvirideae group. The present invention also relates to uses of this polypeptide, in particular for diagnosing disease states and screening for modulator and inhibitor compounds of such polypeptides and in turn the virus itself, screening for infection in vertebrates and biological tissue, cleansing of infected biological tissues, and in the treatment and/or prophylaxis and/or diagnosis of disease caused by a virus of the herpesvirideae group.

Abstract: The present invention provides an improved system for linking nucleic acids to one another. In particular, the present invention provides techniques for producing DNA product molecules that may be easily and directly ligated to recipient molecules. The product molecules need not be cleaved with restriction enzymes in order to undergo such ligation. In preferred embodiments of the invention, the DNA product molecules are produced through iterative DNA synthesis reactions, so that the product molecules are amplified products. The invention further provides methods for directed ligation of product molecules (i.e., for selective ligation of certain molecules within a collection of molecules), and also for methods of exon shuffling, in which multiple different product molecules are produced in a single ligation reaction. Preferred embodiments of the invention involve ligation of product molecules encoding functional protein domains, particularly domains naturally found in conserved gene families.

Abstract: The present invention relates to at least one nucleic acid (i) comprising or consisting of or (ii) encoding a nucleic acid comprising or consisting of, a sequence selected from the group consisting of: 1) SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, and 2) a sequence derived from SEQ ID NO: 1 to 4 by substitution deletion or insertion of at least one nucleotide, provided that a nucleic acid consisting of the sequence derived from SEQ ID NO: 1 to 4 is liable to induce HIV-1 expression in latent HIV-1-infected cells, for use as a medicament, in particular for treating retrovirus infections.

Abstract: The present invention relates to a method for producing a modified foreign chromosome(s) or a fragment(s) thereof, which comprises the steps of: (a) preparing a microcell comprising a foreign chromosome(s) or a fragment(s) thereof, and transferring said foreign chromosome(s) or a fragment(s) into a cell with high homologous recombination efficiency through its fusion with said microcell; (b) in said cell with high homologous recombination efficiency, inserting a targeting vector by homologous recombination into a desired site of said foreign chromosome(s) or a fragment(s) thereof, and/or a desired site of a chromosome(s) derived from said cell with high homologous recombination efficiency, thereby marking said desired site; and (c) in said cell with high homologous recombination efficiency, causing deletion and/or translocation to occur at the marked site of said foreign chromosome(s) or a fragment(s) thereof.

Abstract: HBsAg is expressed in a Saccharomyces cerevisiae host, carrying a plasmid having a HBsAg coding sequence, wherein the plasmid includes: (1) an upstream promoter from a glyceraldehyde-3-phosphate dehydrogenase gene, for controlling expression of the HBsAg coding sequence; and (2) an ARG3 transcription terminator downstream of the HBsAg coding sequence. The plasmids may also include: (3) a LEU2 selection marker; (4) a 2? plasmid sequence; and (5) an origin of replication functional in Escherichia coli. HBsAg can be expressed in this host, and can be purified for use in the manufacture of vaccines, and in particular for the manufacture of combination vaccines and in new monovalent HBV vaccines e.g. with non-alum adjuvants.

Abstract: The invention provides methods and compositions for producing and using minicircle DNA molecules that are useful for plant transformation. The invention also provides methods for transforming plant cells and plants with such minicircle DNA molecules, plant cells and plants produced by such methods, and plants transformed with minicircle DNA molecules. The methods and compositions of the invention are particularly useful for producing “intragenic plants” which do not contain any non-native DNA.

Abstract: In the absence of substantial sequence overlap between a recombinant adenoviral vector and the genome of a packaging cell, helper-dependent E1-containing particles (HDEP) can be formed at low frequency. Provided are means and methods for reducing or preventing the generation of HDEP. To this purpose, novel packaging cells and methods of making these are provided.

Abstract: Disclosed herein are new defective Sindbis viral vectors made from a novel Helper plasmid, with differences in envelope proteins between JT vectors and consensus Sindbis virus sequences, and also between JT and Ar-339 vectors. Also disclosed are vectors produced using the plasmid, methods for producing the vectors, methods for treating mammals suffering from tumors and pharmaceutical formulations for use in the treatment methods.

Abstract: The present invention relates to a method for producing herpes simplex virus (HSV) amplicon particles which includes co-transfecting a host cell with the following: (i) an amplicon vector comprising an HSV origin of replication, an HSV cleavage/packaging signal, and a heterologous transgene expressible in a patient, (ii) one or more vectors individually or collectively encoding all essential HSV genes but excluding all cleavage/packaging signals, and (iii) a vhs expression vector encoding a virion host shutoff protein; and then isolating HSV amplicon particles produced by the host cell, the HSV amplicon particles including the transgene. Also disclosed are a system and a kit for preparing HSV amplicon particles, HSV amplicon particles prepared according to the process of the present invention, and their use.

Abstract: The present invention provides reagents and methods for inhibiting bacterial infection and abnormal cell growth, as well as for selection cloning of nucleic acid inserts.

Abstract: Nucleic acid constructs containing HIV-1 gag/pol and SIV gag or SIV env genes which have been mutated to remove or reduce inhibitory/instability sequences are disclosed. Viral particles and host cells containing these constructs and/or viral particles are also disclosed. The exemplified constructs and viral particles of the invention may be useful in gene therapy for numerous disorders, including HIV infection, or as a vaccine for HIV-1 immunotherapy and immunoprophylaxis.

Abstract: The present invention concerns methods and compositions for in vivo and in vitro targeting. A large number of targeting peptides directed towards human organs, tissues or cell types are disclosed. The peptides are of use for targeted delivery of therapeutic agents, including but not limited to gene therapy vectors. A novel class of gene therapy vectors is disclosed. Certain of the disclosed peptides have therapeutic use for inhibiting angiogenesis, inhibiting tumor growth, inducing apoptosis, inhibiting pregnancy or inducing weight loss. Methods of identifying novel targeting peptides in humans, as well as identifying endogenous receptor-ligand pairs are disclosed. Methods of identifying novel infectious agents that are causal for human disease states are also disclosed. A novel mechanism for inducing apoptosis is further disclosed.

Abstract: The invention provides compositions, methods, and kits for increasing transport of agents across the blood brain barrier while allowing their activity once across the barrier to remain substantially intact. The agents are transported across the blood brain barrier via one or more endogenous receptor-mediated transport systems. In some embodiments the agents are therapeutic, diagnostic, or research agents.

Abstract: Host vector system and methods for plasmid DNA and recombinant protein production. The system allows copy number control of a ColE1 plasmid in E. coli by an RNA molecule that is transcribed from the host's genome and that interacts with plasmid-transcribed RNAI or RNAII. The system can be extended to combine PCN control and antibiotic-free selection.

Abstract: The present invention relates to permanent cell lines from chiropterans suitable for amplification and production microbial agents, preferably viruses, and its use for diagnostic or therapeutic purposes.

Abstract: The present invention relates to novel polypeptides according to caroase 01-05 or any functional equivalents of any of them, suitable for use in a method for preparing a food products having increased whiteness, the use of the enzyme to increase whiteness of at least part of a food product, a process for preparing a food product wherein the enzyme is used and the food product obtained.

Abstract: Aspects of the invention relate to reconfigurable chassis that allow for rapid construction and optimization of biocircuits.

Abstract: The present invention relates to the cloning and characterization of a prokaryotic DNA repair ligase, which is shown to possess a range of activities that allow the ligation and repair of non-compatible DNA ends and double strand breaks (DSBs). The enzyme has a range of applications in the manipulation and cloning of nucleic acids.

Abstract: Provided is a method of preparing a proliferation-regulated recombinant adenoviral vector effectively, comprising preparing a proliferation-regulated vector plasmid by preparing a restriction enzyme-recognizing sequence in a vector plasmid having a proliferation-regulating unit and having an E1A region, a protein-coding region in a E1B region, a poly(A) signal sequence, and a recombinase-recognizing sequence in that order from upstream, by deleting an endogenous promoter in the E1A region or an endogenous promoter regulating expression of the protein-coding gene in one protein-coding region of the E1B region thereof and inserting the restriction enzyme-recognizing sequences in the deficient site, and introducing a promoter expressing specifically in a target organ in the restriction enzyme-recognizing sequence in the restriction enzyme-recognizing sequence; and additionally, integrating the proliferation-regulated vector plasmid into a vector plasmid having a adenoviral genome prepared by deleting the E1 regi