Abstract: Materials and methods for using polypeptides containing fragments and variants of endostatin to treat fibrosis are described herein.

Abstract: Materials and methods for using polypeptides containing fragments and variants of endostatin to treat fibrosis are described herein.

Abstract: This document provides materials and methods for producing endostatin fusion polypeptides having anti-fibrotic activity. For example, provided herein are fusion polypeptides having an IgG Fc domain and a portion of human endostatin that can form high molecular weight multimers, as well as methods for producing such fusion polypeptides in plant cells.

Abstract: Materials and methods for using polypeptides containing fragments and variants of endostatin to treat fibrosis are described herein.

Abstract: Methods to derive novel hybrid type 1 interferons that are broadly active against highly pathogenic viruses of biodefense significance are described. Libraries of hybrid interferon genes were produced using gene shuffling, the proteins were expressed, and screened for activity against viruses of interest. Sequences of several broadly active hybrid interferons are described.

Abstract: Described is a method of screening libraries of variant proteins produced in plant leaves using a plant viral vector to identify a gene of interest comprising, inoculating leaves with a library of viruses expressing variant genes, allowing time for infected foci to form, harvesting a leaf, sticking one face of the leaf to a sticky support material to immobilize the leaf and leaving the opposing face of the leaf exposed, abrading the exposed face with granular material, placing the abraded face in contact with a blot membrane having a backing comprising blotting paper, placing the assembly into a vacuum seal bag; evacuating and sealing the bag; removing the assembly and separating the membrane, performing an assay on the membrane to identify an infected focus of interest; recovering virus corresponding to the infected focus; recovering nucleic acid from the virus, and identifying the gene of interest from the nucleic acid.

Abstract: We have generated virus-like particles (VLPs) that can display other proteins through covalent protein-protein linkages mediated by the ‘Dock and Lock’ interaction between the Drosophila NorpA protein and the C-terminal pentapeptide tail of the InaD protein. This interaction may also be mediated by a portion of the SITAC protein and the Tetraspanin L6 Antigen protein. This system can be used to generate high-density scaffolded arrays of epitopes for immunization. This technology can streamline VLP vaccine candidate production, making it possible to rapidly evaluate panels of candidates in response to current vaccine needs and emerging pathogen threats.

Abstract: Described is a method of screening libraries of variant proteins produced in plant leaves using a plant viral vector to identify a gene of interest comprising, inoculating leaves with a library of viruses expressing variant genes, allowing time for infected foci to form, harvesting a leaf, sticking one face of the leaf to a sticky support material to immobilize the leaf and leaving the opposing face of the leaf exposed, abrading the exposed face with granular material, placing the abraded face in contact with a blot membrane having a backing comprising blotting paper, placing the assembly into a vacuum seal bag; evacuating and sealing the bag; removing the assembly and separating the membrane, performing an assay on the membrane to identify an infected focus of interest; recovering virus corresponding to the infected focus; recovering nucleic acid from the virus, and identifying the gene of interest from the nucleic acid.

Abstract: Described is a method of screening libraries of variant proteins produced in plant leaves using a plant viral vector to identify a gene of interest comprising, inoculating leaves with a library of viruses expressing variant genes, allowing time for infected foci to form, harvesting a leaf, sticking one face of the leaf to a sticky support material to immobilize the leaf and leaving the opposing face of the leaf exposed, abrading the exposed face with granular material, placing the abraded face in contact with a blot membrane having a backing comprising blotting paper, placing the assembly into a vacuum seal bag; evacuating and sealing the bag; removing the assembly and separating the membrane, performing an assay on the membrane to identify an infected focus of interest; recovering virus corresponding to the infected focus; recovering nucleic acid from the virus, and identifying the gene of interest from the nucleic acid.

Abstract: Described is a method of screening libraries of variant proteins produced in plant leaves using a plant viral vector to identify a gene of interest comprising, inoculating leaves with a library of viruses expressing variant genes, allowing time for infected foci to form, harvesting a leaf, sticking one face of the leaf to a sticky support material to immobilize the leaf and leaving the opposing face of the leaf exposed, abrading the exposed face with granular material, placing the abraded face in contact with a blot membrane having a backing comprising blotting paper, placing the assembly into a vacuum seal bag; evacuating and sealing the bag; removing the assembly and separating the membrane, performing an assay on the membrane to identify an infected focus of interest; recovering virus corresponding to the infected focus; recovering nucleic acid from the virus, and identifying the gene of interest from the nucleic acid.

Abstract: Methods to derive novel hybrid type 1 interferons that are broadly active against highly pathogenic viruses of biodefense significance are described. Libraries of hybrid interferon genes were produced using gene shuffling, the proteins were expressed, and screened for activity against viruses of interest. Sequences of several broadly active hybrid interferons are described.

Abstract: Herein is described a modified viral vector comprising: a coat protein modified, for example by the addition of a cysteine residue, such that the modified viral vector yields less soluble virus relative to that from an unmodified viral vector upon extraction of plant material infected with the modified viral vector, thereby facilitating purification of a recombinant protein expressed from the modified viral vector. Also described is a method of reducing viral coat protein impurities during purification of a recombinant protein, a method of biocontainment for a recombinant viral vector, and a method of generating virus inoculum for the modified viral vector.

Abstract: The present invention relates to codon optimization utilizing DNA shuffling. A method of producing gene sequences optimized for a desired functional property is described involving synthesizing a library of parental codon variant genes encoding some or all codon choices at some or all amino acid positions of a gene, reassorting the variant codons among the parental codon variant genes using DNA shuffling thereby forming progeny codon variant genes, expressing the progeny codon variant genes in a host; and screening or selecting for progeny codon variant genes encoding a desired functional property.

Abstract: Herein is described a system to combat poxvirus infection wherein antagonists are developed that bind the soluble cytokine receptor but have no significant biological activity in the host, effectively blocking the virus-mediated suppressor of interferon function, thereby permitting the host's own cytokines to stimulate an antiviral response. Alternatively, interferon molecules can be developed that retain biological activity on their native receptors but fail to bind the viral cytokine binding protein, thereby circumventing this virus immune modulation mechanism.

Abstract: We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3? to 5? exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.

Abstract: We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3? to 5? exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. The parent polynucleotides need not be cleaved into fragments prior to annealing heterologous strands. Therefore, no reassembly is required.

Abstract: Herein is described a system to combat poxvirus infection wherein antagonists are developed that bind the soluble cytokine receptor but have no significant biological activity in the host, effectively blocking the virus-mediated suppressor of interferon function, thereby permitting the host's own cytokines to stimulate an antiviral response. Alternatively, interferon molecules can be developed that retain biological activity on their native receptors but fail to bind the viral cytokine binding protein, thereby circumventing this virus immune modulation mechanism.

Abstract: Herein is described a modified viral vector comprising: a coat protein modified, for example by the addition of a cysteine residue, such that the modified viral vector yields less soluble virus relative to that from an unmodified viral vector upon extraction of plant material infected with the modified viral vector, thereby facilitating purification of a recombinant protein expressed from the modified viral vector. Also described is a method of reducing viral coat protein impurities during purification of a recombinant protein, a method of biocontainment for a recombinant viral vector, and a method of generating virus inoculum for the modified viral vector.

Abstract: We describe here an in vitro method of redistributing sequence variations between non-identical polynucleotide sequences, by making a heteroduplex polynucleotide from two non-identical polynucleotides; introducing a nick in one strand at or near a base pair mismatch site; removing mismatched base(s) from the mismatch site where the nick occurred; and using the opposite strand as template to replace the removed base(s) with bases that complement base(s) in the first strand. By this method, information is transferred from one strand to the other at sites of mismatch.

Abstract: We describe here an in vitro method of increasing complementarity in a heteroduplex polynucleotide sequence. The method uses annealing of opposite strands to form a polynucleotide duplex with mismatches. The heteroduplex polynucleotide is combined with an effective amount of enzymes having strand cleavage activity, 3? to 5? exonuclease activity, and polymerase activity, and allowing sufficient time for the percentage of complementarity to be increased within the heteroduplex. Not all heteroduplex polynucleotides will necessarily have all mismatches resolved to complementarity. The resulting polynucleotide is optionally ligated. Several variant polynucleotides result. At sites where either of the opposite strands has templated recoding in the other strand, the resulting percent complementarity of the heteroduplex polynucleotide sequence is increased. Also described are mismatch endonucleases suitable for use in the process.