公开(公告)号：US06833135B1

公开(公告)日：2004-12-21

申请号：US09269717

申请日：1999-06-03

申请人： Jose A. Frazao Moniz Pereira ,  Alcino Freitas Vieira ,  Elsa M. Ribeiro Dos Santos-Anes ,  Miguel A. Da Costa Garcia ,  Paulo J. Da Silva Alves

发明人： Jose A. Frazao Moniz Pereira ,  Alcino Freitas Vieira ,  Elsa M. Ribeiro Dos Santos-Anes ,  Miguel A. Da Costa Garcia ,  Paulo J. Da Silva Alves

IPC分类号： A61K3904

CPC分类号： C12N9/22 ,  A61K39/00 ,  C12N15/74 ,  C12N15/90 ,  C12N2310/111 ,  C12N2799/021 ,  Y10S435/866

摘要： The present invention discloses a DNA fragment that can direct the insertion of heterologous DNA into a specific site (tRNAAla gene) of Mycobacterium spp. genome. This DNA fragment includes the attachment site region (attP) and the integrase gene of the mycobacteriophage Ms6. Heterologous DNA linked to this DNA fragment can be carried into the mycobacterial genome through a site-specific integration mechanism. A new DNA integration process is disclosed that consists in providing the integrase gene on a suicide vector, that transiently produces the integrase required for the insertion of the gene or genes linked to the Ms6 attP region contained in a separated plasmid vector. This process allows the construction of highly stable recombinant mycobacteria which may be used as vaccines and/or therapeutic vehicles.

摘要翻译： 本发明公开了可将异源DNA插入分枝杆菌的特定位点（tRNA 基因）的DNA片段。 基因组 该DNA片段包括附着位点区域（attP）和分枝杆菌噬菌体Ms6的整合酶基因。 与该DNA片段连接的异源DNA可以通过位点特异性整合机制携带到分枝杆菌基因组中。 公开了一种新的DNA整合方法，其包括在自杀载体上提供整合酶基因，其瞬时产生插入分离的质粒载体中包含的Ms6 attP区的基因或基因所需的整合酶。 该方法允许构建可用作疫苗和/或治疗载体的高度稳定的重组分枝杆菌。