1887

Abstract

, a Gram-negative obligate anaerobe and the causative agent of ovine footrot, secretes a number of extracellular proteases, one of which is highly basic in nature. The gene () encoding this basic protease, from virulent strain 198, has been cloned and sequenced. Clone pBR3KB contained the complete gene which constitutively expressed an active protease using its own promoter, when cloned in . However, levels of protease expression were low and unstable when the clone was expressed in liquid culture. A range of strains were examined for stable expression; strains NH274 and SURE were found to be better hosts for stable expression than other commonly used host strains. Stabilization and enhancement of expression was achieved by deletion of the native promoter region and expression from plasmid promoter or promoters, and by modification of culture conditions. The recombinant protease obtained from was indistinguishable from the native enzyme in size, activity, isoelectric point and immunological properties.

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1994-08-01
2021-10-18
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