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1. 发明公开

EP1194165A1 TAGGED EPITOPE PROTEIN TRANSPOSABLE ELEMENT 审中-公开
标题翻译：转座元件，表位标记的蛋白ENCODED
公开(公告)号：EP1194165A1
公开(公告)日：2002-04-10
申请号：EP00937880.3
申请日：2000-05-26
申请人： Oregon Health Sciences University
发明人： HEFFRON, Fred, L. , PARKER, David, C. , ELLEFSON, Dolph, D.
IPC分类号： A61K39/04 , A61K39/112 , A61K39/12 , A61K39/02
CPC分类号： C07K14/70539 , A61K39/0011 , A61K39/12 , A61K39/21 , C12N15/1034 , C12N15/63 , C12N2740/16034 , C12Q1/689 , Y02A50/412
摘要： 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 bindind agent that recognizes the MHC epitope, identifying labeled eukaryotic cells are identified and lysed to externalize 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.

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