Positive selection vector for bacillus sp.

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/74 (2006.01) C12N 1/21 (2006.01) C12N 9/10 (2006.01) C12N 15/75 (2006.01)

Patent

CA 2234876

A positive selection vector is provided useful for the transformation and screening of Gram positive bacteria and particularly Bacillus sp. for the presence of foreign DNA. The vector comprises a mutant gene encoding a signal peptide processing mutation. Expression of the mutant gene in a Bacillus host cell which lacks the ability to metabolize sucrose is lethal when cells are grown in the presence of sucrose. Foreign DNA may be inserted into the vector so as to inactivate the mutant gene thereby permitting the cells to grow in the presence of sucrose and allowing for facile selection of transformants.

L'invention concerne un vecteur de sélection positive, utile pour la transformation de bactéries à Gram positif, en particulier de Bacillus sp., et leur criblage en fonction de la présence d'ADN étranger. Ledit vecteur comprend un gène mutant codant une mutation de maturation du peptide signal. L'expression du gène mutant dans une cellule hôte de Bacillus, laquelle ne présente pas l'aptitude à la métabolisation du sucrose, est létale lorsque des cellules sont développées en présence de sucrose. Un ADN étranger peut être introduit dans le vecteur de façon à inactiver le gène mutant, ce qui permet aux cellules de croître en présence de sucrose et ainsi une sélection facile des transformants.

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