Mycobacteria functional screening and/or expression vectors

Details Mycobacteria functional screening and/or expression vectors Mycobacteria functional screening and/or expression vectors

C - Chemistry, Metallurgy

12

N

C12N 15/74 (2006.01) A61K 35/74 (2006.01) A61K 39/04 (2006.01) C07H 21/00 (2006.01) C07K 14/35 (2006.01) C07K 16/12 (2006.01) C12N 1/21 (2006.01) C12N 15/31 (2006.01) C12N 15/62 (2006.01) C12Q 1/68 (2006.01) G01N 33/569 (2006.01) A61K 39/00 (2006.01)

Patent

CA 2197717

Recombinant screening, cloning and/or expression vector characterized in that it replicates in mycobacteria and contains 1) a mycobacteria functional replicon; 2) a selection marker, 3) a reporter cassette comprising a) a multiple cloning site (polylinker) b) a transcription terminator which is active in mycobacteria and is located upstream of the polylinker, and c) a coding nucleotide sequence derived from a gene coding for an expression, export and/or secretion protein marker, the nucleotide sequence being deprived of its initiation codon and its regulating sequences. This vector is used for identification and expression of exporter polypeptides, such as the Mycobacterium tuberculosis P28 antigen.

L'invention concerne un vecteur recombinant de criblage et/ou de clonage et/ou d'expression, caractérisé en ce qu'il se réplique chez des mycobactéries, en ce qu'il contient: 1) un réplicon fonctionnel chez les mycobactéries; 2) un marqueur de sélection; 3) une cassette reporteur comprenant a) un site de clonage multiple (polylinker), b) un terminateur de transcription actif chez les mycobactéries, situé en amont du polylinker, et c) une séquence nucléotidique codante issue d'un gène codant pour un marqueur d'expression, et/ou d'exportation et/ou de sécrétion de protéine, ladite séquence nucléotidique étant dépourvue de son codon d'initiation et de ses séquences de régulation. Ce vecteur est utilisé pour l'identification et l'expression de polypeptides exportés, tel que l'antigène P28 de Mycobacterium tuberculosis.

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