Abstract: In the present invention parapox genomes were screened using a novel expression strategy to test genes for dendritic cell recruitment activity. One gene was identified, designated as B2WL, that induces dendritic cell accumulation when expressed in skin. In additional testing a second gene, PP30, was identified that exhibited adjuvant activity in the absence of stimulating dendritic cell accumulation at the site of inoculation. When co-inoculated with an antigen-encoding plasmid, both genes acted as adjuvants in stimulating an antibody response to antigens. Furthermore, nucleic acids encoding the identified B2WL peptide adjuvant enhanced the level of protection against viral infection provided by immunization with an HA-expression plasmid. Thus, novel adjuvants for genetic immunization are identified.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: The Parapox B2L virus envelope protein is used as an adjuvant to enhance a subject's response to an administered antigen. Both antibody and cellular immune responses can be modified. B2L protein is particularly useful as an adjuvant for poorly immunogenic tumor vaccines and subunit vaccines, such as those useful for preventing and/or treating flu, tuberculosis, respiratory syncytial virus, anthrax and HIV.

Abstract: The Parapox B2L virus envelope protein is used as an adjuvant to enhance a subject's response to an administered antigen. Both antibody and cellular immune responses can be modified. B2L protein is particularly useful as an adjuvant for poorly immunogenic tumor vaccines and subunit vaccines, such as those useful for preventing and/or treating flu, tuberculosis, respiratory syncytial virus, anthrax and HIV.

Abstract: The Parapox B2L virus envelope protein is used as an adjuvant to enhance a subject's response to an administered antigen. Both antibody and cellular immune responses can be modified. B2L protein is particularly useful as an adjuvant for poorly immunogenic tumor vaccines and subunit vaccines, such as those useful for preventing and/or treating flu, tuberculosis, respiratory syncytial virus, anthrax and HIV.

Abstract: In the present invention parapox genomes were screened using a novel expression strategy to test genes for dendritic cell recruitment activity. One gene was identified, designated as B2WL, that induces dendritic cell accumulation when expressed in skin. In additional testing a second gene, PP30, was identified that exhibited adjuvant activity in the absence of stimulating dendritic cell accumulation at the site of inoculation. When co-inoculated with an antigen-encoding plasmid, both genes acted as adjuvants in stimulating an antibody response to antigens. Furthermore, nucleic acids encoding the identified B2WL peptide adjuvant enhanced the level of protection against viral infection provided by immunization with an HA-expression plasmid. Thus, novel adjuvants for genetic immunization are identified.

Abstract: The instant invention relates to antigens and nucleic acids encoding such antigens obtainable by screening the Chlamydia psittaci genome. In more specific aspects, the invention relates to methods of isolating such antigens and nucleic acids and to methods of using such isolated antigens for producing immune responses in bovines or other non-human animals. The ability of an antigen to produce an immune response may be employed in vaccination of bovines or antibody preparation techniques.

Abstract: The present application describes novel uses of ruthenium bipyridyls or palladium porphyrins as photo-activatable crosslinking agents. Crosslinking can be between any two molecules including peptides, proteins, or compounds. Crosslinking occurs in the presence of an electron donor such as ammonium persulfate, and requires only moderate intensity visible light. Crosslinking can be between peptides, polypeptides or lead candidate compounds to unknown target molecules. Reagents utilyzing ruthenium bipyridyls and palladium porphyrins crosslinkers for use in diagnostic and detection scenarios are also disclosed.

Abstract: A high throughput oligonucleotide synthesizer is described that includes masks for selectively deblocking of oligonucleotide synthesis sites and the simultaneous addition of reagents to all wells of the plate. The synthesizer includes a multi-well plate, each well of which contains a substrate for oligonucleotide synthesis. The use of masks expedites oligonucleotide synthesis by allowing for rapid delivery of reagents to all wells simultaneously.

Abstract: The instant invention relates to antigens and nucleic acids encoding such antigens obtainable by screening a Chlamydia genome. In more specific aspects, the invention relates to methods of isolating such antigens and nucleic acids and to methods of using such isolated antigens for producing immune responses. The ability of an antigen to produce an immune response may be employed in vaccination or antibody preparation techniques.

Abstract: A method for conferring resistance to a parasite to a host of the parasite, which comprises isolating a gene fragment from the parasite and inserting the gene fragment or a DNA or RNA segment substantially homologous to the gene fragment or to a DNA or RNA sequence functionally equivalent to the gene fragment into the host, wherein (1) transcription of the gene fragment or the DNA or RNA segment in the host occurs in an anti-sense direction, (2) the gene fragment or the DNA or RNA segment is expressed as a gene product in the host, wherein the gene product is capable of disrupting an essential activity of the parasite, or (3) the gene fragment or the DNA or RNA segment is a binding site capable of competing with a native binding site in the parasite, is disclosed along with hosts produced by this process. Particularly preferred is conferring resistance using a gene fragment from a replicase gene of an RNA virus.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.

Abstract: A cooling fan system includes an engine driven mechanical fan and an electrically driven fan. The mechanical fan and the electrical fans are placed in series within a common shroud that directs air between them. In one application, the electrical fan generates an axially directed air flow and the mechanical fan produces a radially directed air flow. The electrical fan is rotated counter to the mechanical fan.

Abstract: A general method for vaccinating against any pathogen is presented. The method utilizes expression library immunization, where an animal is inoculated with an expression library constructed from fragmented genomic DNA of the pathogen. All potential epitopes of the pathogen's proteins are encoded in its DNA, and genetic immunization is used to directly introduce one or more expression library clones to the immune system, producing an immune response to the encoded protein. Inoculation of expression libraries representing portions of the Mycoplasma pulmonis genome was shown to protect mice from subsequent challenge by this natural pathogen. Protection against Listeria.

Abstract: A method for conferring resistance to a parasite to a host of the parasite, which comprises isolating a gene fragment from the parasite and inserting the gene fragment or a DNA or RNA segment substantially homologous to the gene fragment or to a DNA or RNA sequence functionally equivalent to the gene fragment into the host, wherein (1) transcription of the gene fragment or the DNA or RNA segment in the host occurs in an anti-sense direction, (2) the gene fragment or the DNA or RNA segment is expressed as a gene product in the host, wherein the gene product is capable of disrupting an essential activity of the parasite, or (3) the gene fragment or the DNA or RNA segment is a binding site capable of competing with a native binding site in the parasite, is disclosed along with hosts produced by this process. Particularly preferred is conferring resistance using a gene fragment from a replicase gene of an RNA virus.

Abstract: A general method for vaccinating against any pathogen is presented. The method utilizes expression library immunization, where an animal is inoculated with an expression library constructed from fragmented genomic DNA of the pathogen. All potential epitopes of the pathogen's proteins are encoded in its DNA, and genetic immunization is used to directly introduce one or more expression library clones to the immune system, producing an immune response to the encoded protein. Inoculation of expression libraries representing portions of the Mycoplasma pulmonis genome was shown to protect mice from subsequent challenge by this natural pathogen. Protection against Listeria was also obtained using the method.

Abstract: A method for conferring resistance to a parasite to a host of the parasite, which comprises isolating a gene fragment from the parasite and inserting the gene fragment or a DNA or RNA segment substantially homologous to the gene fragment or to a DNA or RNA sequence functionally equivalent to the gene fragment into the host, wherein (1) transcription of the gene fragment or the DNA or RNA segment in the host occurs in an anti-sense direction, (2) the gene fragment or the DNA or RNA segment is expressed as a gene product in the host, wherein the gene product is capable of disrupting an essential activity of the parasite, or (3) the gene fragment or the DNA or RNA segment is a binding site capable of competing with a native binding site in the parasite, is disclosed along with hosts produced by this process. Particularly preferred is conferring resistance using a gene fragment from a replicase gene of an RNA virus.

Abstract: Methods for isolation and purification of recombinant proteins are described. Fusion proteins incorporating a cleavage site sensitive to proteolysis by a plant virus proteinase may be cleaved from carrier proteins to provide high yields of protein product. Methods employing a plant virus proteinase to cleave expressed fusion proteins are particularly suitable for obtaining proteolytically sensitive polypeptides in the presence of added cell protease inhibitors. Also disclosed are recombinant vectors useful for overproducing plant virus proteinases in a suitable host.