METHOD FOR THE ISOLATION OF HIGHLY PURIFIED SALMONELLA FLAGELLINS
METHOD FOR THE ISOLATION OF HIGHLY PURIFIED SALMONELLA FLAGELLINS
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DOI：
10.1128/jcm.22.6.1040-1044.1985
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发表时间：
1985-01-01
影响因子：
9.4
通讯作者：
WALKER, RA
中科院分区：
文献类型：
--
作者：
IBRAHIM, GF;FLEET, GH;WALKER, RA
10种不同的沙门氏菌血清型在补充有0.01%酵母提取物的化学成分确定的培养基中生长。通过离心使细胞沉降后，通过在室温下暴露于pH 2 30分钟来分离鞭毛。通过离心除去无鞭毛细胞，并通过高速离心、硫酸铵沉淀和在50，000分子量截留管中透析进一步纯化上清液中的鞭毛蛋白。这10种鞭毛蛋白制品的纯度都很高，用电镜、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和测定以鞭毛蛋白制品为免疫原的兔抗血清的Salmonella H和O凝集滴度证明了这一点。
Ten different Salmonella serotypes were grown in a chemically defined medium supplemented with 0.01% yeast extract. After sedimentation of the cells by centrifugation, flagella were detached by exposure to pH 2 for 30 min at room temperature. The flagellaless cells were removed by centrifugation, and the flagellin in the supernatant was further purified by high-speed centrifugation, ammonium sulfate precipitation, and dialysis in 50,000-molecular-weight-cutoff tubing. The 10 flagellin preparations were of a high degree of purity, as demonstrated by electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and measurement of salmonella H and O agglutination titers of antisera raised in rabbits with the flagellin preparations as immunogens.