Compositions containing sortase-anchored surface proteins of Streptococcus uberis
A technology of Streptococcus uberis and anchoring protein, which can be used in medical preparations containing active ingredients, antibacterial drugs, bacterial antigen components, etc., and can solve problems such as large changes in numbers
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Embodiment 1
[0080] Example 1 : Streptococcus uberis SrtA Mutant preparation and identification
[0081] Methods and Materials
[0082] Strains and Reagents
[0083] S.uberis strain 0140J, originally isolated from a clinical case of bovine mastitis in the United Kingdom, was used throughout the study. Bacteria were routinely grown in Todd Hewitt or Brain Heart Infusion broth.
[0084] Skim milk was prepared from fresh milk aseptically collected from several cows in the dairy herd of the Institute for Animal Health. Milk was collected from animals free of intramammary infection. After centrifugation (3,000 x g, 10 min), skim milk was carefully removed from the upper flat-layer and pellet of sedimented cells. Put 500μl of milk directly on the blood agar plate containing aesculin (1.0%, w / v; ABA) and culture it with 5ml of milk and an equal volume of Todd Hewitt broth. Colonies for sterility testing of skim milk. In both cases, the plates were incubated at 37°C for 18 hours...
Embodiment 2
[0123] Example 2 : An investigation of the requirement for specific sortase-anchored proteins in the virulence of Streptococcus uberis
[0124] Following recognition of the sortase-anchored proteins, each of the genes encoding these proteins was mutated within the wild-type strain. Mutants, each lacking an individual sortase-anchored protein, were used in an immunoassay model of cows to test for toxicity. Proteins deleted from those that exhibit reduced or eliminated toxicity are associated with the pathogenesis / pathology of the disease. Induction of a neutralizing immune (antibody) response to any and preferably all of these proteins will result in less severe disease following infection with the wild-type strain. Therefore, a vaccine containing any or all of those identified as having a role in virulence would help prevent bovine mastitis.
[0125] method
[0126] Lack SrtA ankyrin ( Sub0135, Sub0145, Sub0207, Sub0241, Sub 0826, Sub0888, Sub1095, Sub1154, S...
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