Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Reexamination Certificate
2005-01-25
2005-01-25
Chan, Christina (Department: 1644)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
C435S006120, C435S007200, C435S007310, C435S029000, C435S320100
Reexamination Certificate
active
06846622
ABSTRACT:
A transposable element is provided that as a 3′ and a 5′ end. The transposable element includes a 5′ recombining site 5′ of a nucleic acid sequence encoding a selectable marker, a 3′ recombining site 3′ of the nucleic acid sequence encoding a selectable marker, a nucleic acid sequence encoding and MHC epitope 5′ to the 5′ recombining site or 3′ to the 3′ recombining site, and an insertion end comprising an inverted repeat sequence sufficient for integration of the transposable element at the 5′ and the 3′ end of the transposable element. In one embodiment, a transposable element is provided that has a 5′ and a 3′ end. The transposable element includes a 5′ loxP sequence 5′ of a nucleic acid encoding a selectable marker, a 3′ loxP sequences 3′ of a nucleic acid encoding the selectable marker, an MHC epitope 5′ to the 5′ loxP sequences or 3′ of the 3′ loxP sequence, an insertion end at the 5′ end of the transposable element, and an insertion end at the 3′ of the transposable element. A method is provided for detecting an antigenic epitope of a pathogen by infecting a pathogenic cell with a transposable element of the invention, wherein the infection results in the integration of the transposable element in a nucleic acid sequence of the bacterial cell; transforming the pathogenic cell with a vector comprising a transposase; contacting a eukaryotic cell that can internalize the pathogenic cell with the pathogenic cell infected with the transposable element; contacting the eukaryotic cell with a specific binding partner that recognizes the MHC epitope; identifying the labeled eukaryotic cells and externalizing the bacteria cell. The externalized bacterial cell may be grown to produce a population of bacterial cells, and the nucleic acid sequence of the bacterial cell that has the integrated transposable element is identified. This nucleic acid sequence encodes the antigenic element of the panthogen. A method is also provided for generating a carrier vaccine by infecting a bacterial cell with the transposable element of the invention, wherein the transposable further comprises an antigen associated with a disease operably linked to the MHC epitope of the transposable element. The infection of the bacteria results in the integration of the transposable element in a nucleic acid sequence of the bacterial cell. The pathogenic cell is then internalized into a eukaryotic cell and the eukaryotic cell is exposed to a specific bindned agent that recognizes the MHC epitope, identifying labeled eukaryotic cells are identified and lysed to externalized the bacteria cell, which is cultured to produce a population of bacterial cells. The nucleic acid sequence of the bacterial cell that has the integrated transposable element is identified, wherein the nucleic acid sequence encodes the antigenic element of the pathogen. The graving bacterial cell identified, and may be used as the carrier vaccine.
REFERENCES:
Goldsby, RA et al. Kuby Immunology: 4thEdition. [2000] Goldsby et al, eds. W.H. Freeman and Co., New York. p. 209.*
McMahon et al., “Transposon-Mediated Random Insertions and Site-Directed Mutagenesis Prevent the Trafficking of a Mouse Mammary Tumor Virus Superantigen,”Virology 243:354-365 (1998).
Szalay et al., “Presentation of Lysteria Monocytogenes Antigens by Major Histocompatibility Complex Class I Molecules to CD8 Cytotoxic T Lymphocytes Independent of Lysteriolysin Secretion and Virulence,”Eur. J. Immunol. 24:1471-1477 (1994).
Ellefson Dolph D.
Heffron Fred L.
Parker David C.
Chan Christina
Klarquist & Sparkman, LLP
Oregon Health & Science University
Vandervegt F. Pierre
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