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1. 发明申请

WO2009009798A3 ALLELIC EXCHANGE MUTAGENESIS IN MAP 审中-公开
标题翻译：地图中的等位交换致突变
公开(公告)号：WO2009009798A3
公开(公告)日：2009-07-30
申请号：PCT/US2008069999
申请日：2008-07-14
申请人： UNIV WASHINGTON STATE , DAVIS WILLIAM C , HAMILTON MARY JO , DAHL JOHN , PARK KUN TAEK
发明人： DAVIS WILLIAM C , HAMILTON MARY JO , DAHL JOHN , PARK KUN TAEK
IPC分类号： C12N15/31
CPC分类号： C12N15/102 , A61K39/04 , A61K2039/522 , C12R1/325
摘要： Particular aspects provide efficient allelic exchange methods to generate directed mutations within genes of slow-growing stains of mycobacteria (e.g., Mycobacterium avium subsp. paratuberculosis (Map), Map10 or GFP-expressing Map K-10) using a phage-delivery system, and demonstrate high efficiency allelic exchange. Additional exemplary aspects provide non-naturally occurring slow-growing strains of mycobacteria (e.g., Map, M. bovis, M. tuberculosis) having at least one gene deletion (e.g., pknG, relA, lsr2, panC, panD, proC, trpD, sapM (MAP3432), lysA_1, leuD, and leuC, and deletion mutants at the orthologous loci of two known virulence genes in pathogenic mycobacteria (relA and pknG) and one gene related to colony morphology and biofilm formation in fast growing mycobacteria (lsr2) were made. Further aspects provide novel vaccines comprising such deletion mutants, or portions thereof, and methods for making said vaccines. Yet further aspects provide methods for protecting a mammal from virulent Map, M. bovis, or M. tuberculosis, comprising treating the mammal with the inventive vaccines.
摘要翻译：具体方面提供了使用噬菌体递送系统在缓慢生长的分枝杆菌染色体（例如鸟分枝杆菌副结核亚种（Map），Map10或GFP表达Map K-10）的基因内产生定向突变的有效等位基因交换方法，以及展示高效率的等位基因交换。其他示例性方面提供了具有至少一个基因缺失的非天然存在的慢性分枝杆菌菌株（例如，Map，M.bovis，结核分枝杆菌）（例如pknG，relA，lsr2，panC，panD，proC，trpD，在致病性分枝杆菌中两种已知毒力基因（relA和pknG）的直系同源基因座上的sapM（MAP3432），lysA_1，leuD和leuC以及与快速生长的分枝杆菌（lsr2）中的菌落形态和生物膜形成有关的一种基因的缺失突变体分别是进一步的方面提供了用于保护哺乳动物免受毒性Map，牛分枝杆菌或结核分枝杆菌的方法，所述方法包括用本发明的疫苗。

2. 发明申请

WO2009009798A2 ALLELIC EXCHANGE MUTAGENESIS IN MAP 审中-公开
标题翻译：地图中的ALLELIC EXCHANGE MUTAGENESIS
公开(公告)号：WO2009009798A2
公开(公告)日：2009-01-15
申请号：PCT/US2008/069999
申请日：2008-07-14
申请人： WASHINGTON STATE UNIVERSITY , DAVIS, William C. , HAMILTON, Mary Jo , DAHL, John , PARK, Kun Taek
发明人： DAVIS, William C. , HAMILTON, Mary Jo , DAHL, John , PARK, Kun Taek
IPC分类号： C12N15/09 , C12N1/21
CPC分类号： C12N15/102 , A61K39/04 , A61K2039/522 , C12R1/325
摘要： Particular aspects provide efficient allelic exchange methods to generate directed mutations within genes of slow-growing stains of mycobacteria ( e.g., Mycobacterium avium subsp. paratuberculosis (Map), Map 10 or GFP-expressing Map K-10) using a phage-delivery system, and demonstrate high efficiency allelic exchange. Additional exemplary aspects provide non-naturally occurring slow-growing strains of mycobacteria ( e.g., Map, M. bovis, M. tuberculosis ) having at least one gene deletion ( e.g., pknG, relA, lsr2 , panC, panD, proC, trpD, sapM (MAP3432), lysA_1, leuD , and leuC, and deletion mutants at the orthologous loci of two known virulence genes in pathogenic mycobacteria ( rel A and pkn G) and one gene related to colony morphology and biofilm formation in fast growing mycobacteria (lsr 2) were made. Further aspects provide novel vaccines comprising such deletion mutants, or portions thereof, and methods for making said vaccines. Yet further aspects provide methods for protecting a mammal from virulent Map, M. bovis, or M. tuberculosis , comprising treating the mammal with the inventive vaccines.
摘要翻译：具体方面提供了有效的等位基因交换方法，以使用噬菌体递送系统在分枝杆菌慢生长染色基因（例如鸟分枝杆菌亚型副结核（Map），Map10或GFP表达的Map K-10）的基因内产生定向突变，以及表明高效等位基因交换。另外的示例性方面提供具有至少一个基因缺失的非天然存在的慢生长的分枝杆菌菌株（例如，Map，M.bovis，M.tuberculosis）（例如，pknG，relA，lsr2，panC，panD，proC，trpD， sapM（MAP3432），lysA_1，leuD和leuC，以及病原分枝杆菌（relA和pknG）中两种已知毒力基因的直向同源基因座的缺失突变体，以及与快速生长的分枝杆菌（lsr2）中的集落形态和生物膜形成有关的一个基因进一步的方面提供了包含这种缺失突变体或其部分的新型疫苗以及制备所述疫苗的方法。另外的方面提供了保护哺乳动物免受有毒的地图，牛分枝杆菌或结核分枝杆菌的方法，包括用哺乳动物治疗本发明的疫苗。

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