Enhanced expression from the pm promoter

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/09 (2006.01)

Patent

CA 2673215

The present invention concerns a method of producing a desired gene product in a recombinant gene expression system, said method comprising expressing said gene from a Pm promoter-based expression system using at least two mutant elements selected from: (i) a mutant Pm promoter; (ii) a mutant mRNA leader; and (iii) a mutant XyIS; wherein said mutant elements each comprise one or more mutations which enhance expression of said desired gene. Particularly combinations of a mutant Pm promoter and a mutant mRNA leader are concerned. Isolated nucleic acid molecules, vectors, host cells, libraries, expression systems, methods of enhancing expression, obtaining nucleic acid molecules and identifying combination mutants which enhance expression, artificially constructed operons and their uses are also encompassed.

La présente invention concerne un procédé destiné à produire un produit génétique souhaité dans un système d'expression de gène recombinant, ledit procédé consistant à exprimer ledit gène à partir d'un système d'expression basé sur un promoteur Pm, à l'aide d'au moins deux éléments mutants choisis parmi : (i) un promoteur Pm mutant ; (ii) une séquence de tête d'ARNm mutant ; et (iii) un XyIS mutant. Dans ledit procédé, chacun desdits éléments mutants comporte une ou plusieurs mutation(s) qui améliore(nt) l'expression dudit gène souhaité. En particulier, l'invention concerne des combinaisons d'un promoteur Pm mutant et d'une séquence de tête d'ARNm mutant. L'invention concerne également des molécules d'acide nucléique isolées, des vecteurs, des cellules hôtes, des bibliothèques, des systèmes d'expression, des procédés destinés à améliorer l'expression, à obtenir des molécules d'acide nucléique et à identifier des mutants combinés qui améliorent l'expression, des opérons construits artificiellement et leurs utilisations.

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