GENETIC-TRANSFORMATION IN STAPHYLOCOCCUS-AUREUS - ISOLATION AND CHARACTERIZATION OF A COMPETENCE-CONFERRING FACTOR FROM BACTERIOPHAGE-80-ALPHA LYSATES
GENETIC-TRANSFORMATION IN STAPHYLOCOCCUS-AUREUS - ISOLATION AND CHARACTERIZATION OF A COMPETENCE-CONFERRING FACTOR FROM BACTERIOPHAGE-80-ALPHA LYSATES
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DOI：
10.1128/jb.148.1.301-307.1981
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发表时间：
1981-01-01
影响因子：
3.2
通讯作者：
PATTEE, PA
中科院分区：
文献类型：
--
作者：
BIRMINGHAM, VA;PATTEE, PA
葡萄球菌噬菌体80α粗裂解产物中的能力授予活性。经(NH_4)_2SO_4沉淀、差速超速离心法和Ficoll速率区带离心法浓缩纯化。这种浓缩制剂对金黄色葡萄球菌8325-4的测定细胞显示出不能归因于残留的80.α的裂解活性。有感染性颗粒存在。对集中的能力授予活动的EM检查显示，偶尔会有完整但空虚的病毒粒子和大量游离的噬菌体尾巴。对该物质的十二烷基硫酸钠-聚丙烯酰胺凝胶分析证实，赋予活性的能力只包含一些主要病毒粒子蛋白，而不是全部。当检测细胞和80α时，赋予能力的活性呈单击式动力学。DNA的转染率过高。因此，赋予能力的活动似乎是80α的一个独特的形态发生的前身。病毒粒子在0.1M CaCl2存在的情况下介导转染和转化的病毒粒子。
The competence-conferring activity in crude lysates of the staphylococcal bacteriophage 80.alpha. was concentrated and purified by (NH4)2SO4 precipitation, differential ultracentrifugation and rate-zonal centrifugation through Ficoll. This concentrated preparation exhibited lytic activity toward assay cells of S. aureus 8325-4 that could not be attribute to the residual 80.alpha. infectious particles present. EM examination of the concentrated competence-conferring activity revealed an occasional intact but empty virion and large numbers of free phage tails. Sodium dodecyl sulfate-polyacrylamide gel analysis of this material confirmed that the competence-conferring activity contained only some, but not all, of the major virion proteins. The competence-conferring activity exhibited single-hit kinetics when assay cells and 80.alpha. transfecting DNA were present in excess. The competence-conferring activity thus seems to be a unique morphogenic precursor of the 80.alpha. virion that mediates transfection and transformation in the presence of 0.1 M CaCl2.