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首页> 外文期刊>African Journal of Biotechnology >Lysis of mastitis pathogens isolated from dairy cow milk samples by purified recombinant lysostaphin
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Lysis of mastitis pathogens isolated from dairy cow milk samples by purified recombinant lysostaphin

机译：通过纯化的重组溶葡萄球菌素裂解乳牛乳样品中分离出的乳腺炎病原体

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To avoid the use of broad spectrum antibiotics which can induce antibiotic resistance, we developed a recombinant lysostaphin with high purity against mastitis pathogens isolated from dairy cow milk. The gene encoding for mature lysostaphin peptide was cloned into an?Escherichia coli?expression systems, pET28a, to produce recombinant lysostaphin. This recombinant lysostaphin was purified by a His60 Ni Gravity Column to obtain milligram amounts of recombinant lysostaphin protein with high purity. Staphylococcal mastitis pathogens, which were isolated from 30 milk samples from cows diagnosed with mastitis, were identified on the basis of colony characteristics, Gram staining, pigment production, hemolysis on 5% bovine blood agar and tube coagulase test. The antibiotic resistance of the isolates was determined by disc diffusion assay. Most of the tested isolates were resistant to some antibiotics. All the tested strains were lysed by the purified recombinant lysostaphin in turbidity reduction assay. The recombinant lysostaphin was found capable of killing?Staphylococcipathogens?in vitro?within an hour (minimum inhibitory concentration (MIC) 2.5 to 16 μg/ml) and was active in a milk-like environment. Thus, the recombinant lysostaphin from the pET system is effective and is expected to complement clinical treatment of cow mastitis.
机译：为了避免使用可引起抗生素耐药性的广谱抗生素，我们开发了一种高纯度的溶葡萄球菌素，可抗击从乳牛乳中分离出的乳腺炎病原体。将编码成熟溶葡萄球菌素肽的基因克隆到大肠杆菌表达系统pET28a中，以生产重组溶葡萄球菌素。通过His 60 Ni重力柱纯化该重组溶葡萄球菌素，得到毫克量的高纯度的重组溶葡萄球菌素蛋白。从菌落特征，革兰氏染色，色素生成，5％牛血琼脂上的溶血和试管凝结酶试验的基础上，鉴定出葡萄球菌性乳腺炎病原体，这些病原体是从诊断为乳腺炎的母牛的30份牛奶样本中分离出来的。分离物的抗生素抗性通过盘扩散测定法确定。大多数测试分离株对某些抗生素具有抗性。在浊度降低试验中，所有测试的菌株都用纯化的重组溶葡萄球菌素裂解。发现该重组溶葡萄球菌素能够在一个小时内（最小抑菌浓度（MIC）为2.5至16μg/ ml）在一个小时内杀死“葡萄球菌病原体”，并且在类似牛奶的环境中具有活性。因此，来自pET系统的重组溶葡萄球菌素是有效的，并有望补充牛乳腺炎的临床治疗。

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