Novel mutants of gram negative mucosal bacteria and...

C - Chemistry – Metallurgy – 12 – N

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C12N 1/20 (2006.01) A61K 39/02 (2006.01) A61K 39/095 (2006.01) A61K 39/10 (2006.01) A61P 31/04 (2006.01) C07K 14/195 (2006.01) C07K 14/22 (2006.01) C07K 14/235 (2006.01) C07K 14/705 (2006.01) A61K 39/00 (2006.01)

Patent

CA 2300854

It is possible to inactivate the early stage of lipid A synthesis of mucosal gram negative bacteria without compromising cell viability. In particular the lpxA gene in N. meningitidis was mutated and resulting lpxA knockout mutants were found to be completely lipopolysaccharide(LPS)-deficient. The major outer membrane proteins (OMPs) were detected in normal amounts. The finding provides important implications for understanding of structure and biogenesis of the outer membrane. On a practical level, the availability of LPS-deficient mutants of pathogenic mucosal bacteria such as N. meningitidis opens up new avenues to vaccine development. It enables easy isolation of endotoxin-free purified proteins, outer membranes or even whole-cell preparations for use in immunisation.

Il est possible d'inactiver la première étape de la synthèse de lipides A de bactéries de muqueuses Gram négatives sans compromettre la viabilité cellulaire. En particulier, on a fait subir une mutation au gène lpxA dans N. meningitidis et on a découvert que les mutants inactivés de lpxA obtenus étaient complètement déficients en lipopolysaccharides (LPS). On a détecté des quantités normales de protéines principales de membrane externe (OMP) . La présente invention a des implications importantes pour la compréhension de la structure et de la biogenèse de la membrane externe. Sur un plan pratique, la découverte de mutants déficients en LPS de bactéries pathogènes des muqueuses telles que N. meningiditis ouvre de nouvelles perspectives dans le développement de vaccins. Elle permet un isolement facile de protéines purifiées exemptes d'endotoxines, de membranes externes ou même de préparations de cellules entières s'utilisant pour une immunisation.

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