Abstract: The present disclosure concerns methods and compositions for inhibiting replication of viruses in mammalian cells. In some cases the virus can be African Swine Fever virus, or related viruses. The methods described herein can make use of programmable nucleases.

Abstract: Biosecure algae and methods for preparing biosecure algae that have a substantially decreased capability to survive in a natural environment are described. The methods include transforming a genetically modified alga to include an essential gene that is operably linked to a promoter system that is active only in the presence of an inducer compound, transforming the genetically modified alga to include a lethal gene that is operably linked with a promoter system that is inactive only in the presence of a repressor compound. The biosecure algae are only able to survive in an artificial algae culture that includes factors or conditions not found in a natural environment.

Abstract: Infectious pancreatic necrosis virus (IPNV), the etiologic agent of infectious pancreatic necrosis in salmonid fish, causes significant losses to the aquaculture industry. The gene for the viral capsid protein (VP2) was cloned into a yeast expression vector and expressed in Saccharomyces cerevisae. Expression of the capsid gene in yeast resulted in formation of approximately 20 nanometer sub-viral particles composed solely of VP2 protein. Anti-IPNV antibodies were detected in rainbow trout vaccinated either by injection of purified VP2-subviral particles (rVP2-SVP) or by feeding recombinant yeast expressing rVP2-SVP. Challenge of rVP2-SVP immunized trout with a heterologous IPNV strain and subsequent viral load determination showed that both injection and orally vaccinated fish had lower IPNV loads than naive or sham-vaccinated fish.

Abstract: Recombinant oleaginous alga that include one or more heterologous genes that increase the ability of the alga to use one or more natural saccharides such as cellulosic or hemicellulosic sugars for algal growth are described. The recombinant oleaginous algae are transformed to include one or more genes expressing sugar metabolizing enzymes or sugar transporting proteins, along with suitable control elements. Use of natural saccharides as a carbon source can allow the algae to produce biofuel precursors in a relatively efficient manner. Processes for preparing the alga, growing the alga, and extracting the biofuel precursors from the alga are also described.

Abstract: Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization.

Abstract: Methods and constructs for genetic manipulation of one or more of shrimp, shellfish, mollusks, and fish are disclosed. The nucleic acid construct includes a promoter and an internal ribosome entry site of an insect picomavirus, such as a cricket paralysis-like picomavirus. One or more open reading frames can be operably associated with one or both of the promoter and the internal ribosome entry site, and one or more proteins or protein subunits can be expressed upon introduction of the construct into a host cell, such as into a shrimp. Method for producing immortalized crustacean cell lines using enhancer elements derived from shrimp and/or shrimp viruses are also described.

Abstract: Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization.

Abstract: Methods and constructs for genetic manipulation of one or more of shrimp, shellfish, mollusks, and fish are disclosed. The nucleic acid construct includes a promoter and an internal ribosome entry site of an insect picomavirus, such as a cricket paralysis-like picomavirus. One or more open reading frames can be operably associated with one or both of the promoter and the internal ribosome entry site, and one or more proteins or protein subunits can be expressed upon introduction of the construct into a host cell, such as into a shrimp. Method for producing immortalized crustacean cell lines using enhancer elements derived from shrimp and/or shrimp viruses are also described.

Abstract: Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization.

Abstract: A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.

Abstract: Infectious pancreatic necrosis virus (IPNV), the etiologic agent of infectious pancreatic necrosis in salmonid fish, causes significant losses to the aquaculture industry. The gene for the viral capsid protein (VP2) was cloned into a yeast expression vector and expressed in Saccharomyces cerevisae. Expression of the capsid gene in yeast resulted in formation of approximately 20 nanometer sub-viral particles composed solely of VP2 protein. Anti-IPNV antibodies were detected in rainbow trout vaccinated either by injection of purified VP2-subviral particles (rVP2-SVP) or by feeding recombinant yeast expressing rVP2-SVP. Challenge of rVP2-SVP immunized trout with a heterologous IPNV strain and subsequent viral load determination showed that both injection and orally vaccinated fish had lower IPNV loads than naive or sham-vaccinated fish.

Abstract: A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.

Abstract: A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.

Abstract: Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically-derived energy. In this study it is shown that the microalga Phaeodaclylurn tricornutum can be engineered to import glucose and grow in the dark through the introduction of genes encoding glucose transporters. Both the human and Chlorella kessleri glucose transporters facilitated the uptake of glucose by P. tricornutum, allowing the cells to metabolize exogenous organic carbon and thrive, independent of light. This is the first successful trophic conversion of an obligate photoautotroph through metabolic engineering, and it demonstrates that methods of cell nourishment can be fundamentally altered with the introduction of a single gene. Since strains transformed with the glucose transport genes are able to grow non-photosynthetically, they can be exploited for the analysis of photosynthetic processes through mutant generation and characterization.

Abstract: In a method for quantitating an analyte by measuring time resolved transfer of fluorescence energy to or from a label quantitatively associated with the analyte, the present invention provides an improvement comprising measuring the energy transferred from donor compounds having the ability to absorb light energy and then transfer this energy to cross-linked allophycocyanin in a time-resolved manner, where the cross-linked allophycocyanin used according to this invention has not been exposed to strongly chaotropic agents after cross-linking.

Abstract: A crustacean or rotifer is infected with a recombinant infectious virus that expresses a protein exogenous to the virus. The genome of the crustacean or rotifer itself remains unaltered. Crustacean, rotifer, insect, or viral promoters drive the transcription of a gene inserted into the recombinant virus genome, and the virus replicates in the crustacean or rotifer cell cytoplasm. The infected crustacean or rotifer can be provided directly to humans or non-human animals, or, following production and harvest of the crustaceans or rotifers, purified recombinant protein or polypeptide can be provided. Large quantities of biopharmaceuticals can be produced rapidly and inexpensively using this production system.

Abstract: In a method for quantitating an analyte by measuring time resolved transfer of fluorescence energy to or from a label quantitatively associated with the analyte, the present invention provides an improvement comprising measuring the energy transferred from donor compounds having the ability to absorb light energy and then transfer this energy to cross-linked allophycocyanin in a time-resolved manner, where the cross-linked allophycocyanin used according to this invention has not been exposed to strongly chaotropic agents after cross-linking.

Abstract: This invention is directed to the utilization of the developing methods for molecular manipulation of cyanobacteria and red algae (and potentially cryptomonad algae) to express of phycobiliproteins and phycobiliprotein linker fusion proteins and their utilization as phycobiliprotein, phycobilisome and subassembly based reagents. In particular, the present invention relates to a method for a specific binding assay to determine a target moiety which is a member of a specific binding pair, and provides an improvement in the method comprising using a detectable label which is a fusion protein containing both a phycobiliprotein domain and another domain corresponding to a first member of a specific binding pair, where the fusion protein binds to a second member of the specific binding pair to provide a detectable labeled complex.

Abstract: Genetic fusions for use in genetic engineering of eukaryotic algae employ a promoter from a light harvesting protein fused to a protein of interest. The fusions can be introduced and selected using an antibiotic resistance determinant. One gene useful for such selection is the sh ble gene encoding a bleomycin binding protein.

Abstract: Combinatorial libraries of labeled biochemical compounds and methods for producing such combinatorial libraries comprising the steps of producing labeled individual units, combining at least two of the labeled individual units so as to produce a labeled biochemical compound, and repeating this process at least once so as to produce a combinatorial library of labeled biochemical compounds. Also, methods for determining the conformation of a biochemical compound which comprise producing a combinatorial library of labeled biochemical compounds, contacting the combinatorial library of labeled biochemical compounds with a target receptor molecule so that a selected labeled biochemical compound binds to the target receptor molecule, and determining the conformation of the selected labeled biochemical compound when bound to the receptor molecule.