Modified construct downstream of the initiation codon for...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/71 (2006.01) C12N 15/67 (2006.01)

Patent

CA 2413612

The invention concerns a construct for expressing a gene coding for a recombinant protein of interest placed under the control of a tryptophan operon (Ptrp) in a prokaryotic cell, comprising directly downstream of the initiation codon a nucleic sequence SEQ ID N?o¿ 1 and downstream of said sequence a multiple cloning cassette designed to receive the gene coding for said recombinant protein of interest, at least nucleic acids of the nucleic sequence SEQ ID N?o¿ 1 being mutated or deleted so as to enable overexpression of said recombinant protein. The invention also concerns a vector containing such a construct, a prokaryotic host cell transformed by said vector, as well as a method for producing a recombinant protein of interest using the inventive construct.

L'invention concerne une construction pour l'expression d'un gène codant pour une protéine recombinante d'intérêt placé sous le contrôle de l'opéron tryptophane Ptrp dans une cellule hôte procaryote, qui comprend directement en aval du codon d'initiation une séquence nucléique SEQ ID N~1 et en aval de cette séquence une cassette de clonage multiple destinée à recevoir le gène codant pour ladite protéine recombinante d'intérêt, au moins un des acides nucléiques de la séquence nucléique SEQ ID N~1 étant muté ou délété de façon à permettre une surexpression de ladite protéine recombinante. L'invention a encore pour objet un vecteur contenant une telle construction, une cellule hôte procaryote transformée par ledit vecteur, tout comme un procédé de production d'une protéine recombinante d'intérêt utilisant une construction selon l'invention.

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