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奇异变形杆菌用于高频转导链霉素 - 磺胺及链霉素 - 磺胺 - 卡那霉素抗性系统的推导及特性

Derivation and properties of Proteus mirabilis systems for high frequency transduction of streptomycin--sulphonamide and streptomycin-sulphonamide--kanamycin resistances.

作者信息

Coetzee J N

出版信息

J Gen Microbiol. 1976 Sep;96(1):95=107. doi: 10.1099/00221287-96-1-95.

Abstract

Properties of two transducing systems with phages capable of high frequency transduction (HFT) of streptomycin and sulphonamide resistance markers of the V group plasmid R905, and of these markers plus the kanamycin resistance marker derived from a previously described HFT phage 5006MHFTak, are described. Transducing particles of the former phage, named 5006MHFTsus, were detected using the replica-plate technique in an ultraviolet-induced lysate of Proteus mirabilis strain PM5006 transduced to streptomycin and sulphonamide resistance by phage 5006M grown on PM5006 carrying R905. Phage 5006MHFTsusk was also detected by the replica-plate technique in ultraviolet-induced lysates of of phage 5006MHFTsus transductants retransduced to ampicillin and kanamycin resistance by phage 5006MHFTak. Both phages were serologically identical to the parent phage 5006M. Ultraviolet-induced lysates transduced their markers to PM5006 at frequencies of about 5 X 10(-2)/plaque-forming unit adsorbed for both the phages. With phage 5006MHFTsusk, this frequency was increased about 10-fold by simultaneous infection of recipients with homologous non-transducing phage, while phage 5006MHFTsus transductions only underwent a twofold increase. Transductants took about 60 min to express complete resistance to 50 mug streptomycin/ml, and resistance to 1600 mug sulphadiazine/ml was complete within 120 min after phage adsorption. Phage 5006MHFTsusk was slightly more resistant to ultraviolet inaction of its transducing potential and reasons are given for the belief that transductants of both phages are heterogenote-like. Both phage lysates were also capable of generalized transduction and, like previously described HFT phages, lysates transduced the leucine marker at increased frequencies. Using previously described extra- and intra-species phages hosts, it was found that the phages could transduce in single infection and were defective in the lysogenic conversion function as well as in a maturation step. Possible modes of formation of the HFT particles are discussed9

摘要

本文描述了两种转导系统的特性,这两种转导系统利用能够对V组质粒R905的链霉素和磺胺抗性标记进行高频转导(HFT)的噬菌体,以及这些标记加上源自先前描述的HFT噬菌体5006MHFTak的卡那霉素抗性标记。使用复制平板技术在奇异变形杆菌菌株PM5006的紫外线诱导裂解物中检测到前一种噬菌体(命名为5006MHFTsus)的转导颗粒,该菌株通过在携带R905的PM5006上生长的噬菌体5006M转导为对链霉素和磺胺具有抗性。在通过噬菌体5006MHFTak再次转导为对氨苄青霉素和卡那霉素具有抗性的5006MHFTsus转导子的紫外线诱导裂解物中,也通过复制平板技术检测到了噬菌体5006MHFTsusk。两种噬菌体在血清学上都与亲本噬菌体5006M相同。紫外线诱导裂解物以约5×10⁻²/吸附的噬菌斑形成单位的频率将其标记转导至PM5006。对于噬菌体5006MHFTsusk,通过用同源非转导噬菌体同时感染受体,该频率增加了约10倍,而噬菌体5006MHFTsus的转导则仅增加了两倍。转导子大约需要60分钟才能对50μg/ml链霉素表现出完全抗性,并且在噬菌体吸附后120分钟内对1600μg/ml磺胺嘧啶的抗性完全形成。噬菌体5006MHFTsusk对其转导潜力的紫外线失活略具抗性,并给出了认为两种噬菌体的转导子类似异源基因的理由。两种噬菌体裂解物也都能够进行普遍性转导,并且与先前描述的HFT噬菌体一样,裂解物以增加的频率转导亮氨酸标记。使用先前描述的种间和种内噬菌体宿主,发现这些噬菌体能够在单次感染中转导,并且在溶原性转化功能以及成熟步骤中存在缺陷。讨论了HFT颗粒的可能形成方式。

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