Plasmid shuttle vector between escherichia coli and bacteria...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/74 (2006.01) C12N 1/21 (2006.01) C12N 15/70 (2006.01) C12N 15/77 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2438118

A plasmid shuttle vector characterized by having a DNA sequence encoding a sequence which is replicable in both of a bacterium belonging to the genus Brevibacillus and Escherichia coli and capable of serving as a promoter in the brevibacillus and as a selection marker for E.coli and the brevibacillus; and a method of transforming a brevibacillus by using the above plasmid shuttle vector. The above plasmid shuttle vector enables efficient transformation of a brevibacillus. This plasmid shuttle vector pNCMO2 has a gene expression regulatory domain whereby a target gene is efficiently secreted and expressed in the brevibacillus but the expression thereof is remarkably inhibited in E. coli.Thus, this plasmid shuttle vector makes it possible to efficiently construct a recombinant gene expression plasmid by using E. coli and is appropriate for the production of a protein in a brevibacillus.

Vecteur plasmide navette caractérisé en ce qu'il possède un ADN codant une séquence qui est réplicable dans une bactérie qui appartient au genre Brevibacillus et Escherichia coli et qui est capable de servir de promoteur dans Brevibacillus ainsi que de marqueur de sélection pour E. coli et Brevibacillus ; et procédé pour transformer Brevibacillus en utilisant le vecteur plasmide navette. Ce vecteur plasmide navette permet la transformation efficace de Brevibacillus. Le vecteur plasmide navette, pNCMO2, comprend un domaine régulateur d'expression génique sécrété et exprimé dans Brevibacillus ; son expression est cependant remarquablement inhibée dans E. coli. De cette manière, le vecteur plasmide navette permet de construire efficacement un plasmide d'expression génique recombinant en utilisant E. coli et s'avère efficace pour la production d'une protéine dans Brevibacillus.

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