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嗜菌体蛭弧菌在热处理大肠杆菌上的周质内生长

Intraperiplasmic growth of Bdellovibrio bacteriovorus on heat-treated Escherichia coli.

作者信息

Hespell R B

出版信息

J Bacteriol. 1978 Mar;133(3):1156-62. doi: 10.1128/jb.133.3.1156-1162.1978.

Abstract

Heat treatment (55 degrees C for 40 min) of cell suspensions in buffer (ca. 3 x 10(9) cells per ml) of Escherichia coli ML35 caused a 4- to 4.5-log loss of cell viability. Similar results were found for several other E. coli strains that were examined. As a result of this heat treatment, 260-nm- and 280-nm-absorbing materials were released into the suspending buffer, along with about 10% of the total cellular radioactivity, when cells uniformly labeled with (14)C were used. In comparison with untreated cells, heat-treated E. coli ML35 cells showed (i) no significant changes in macromolecular composition other than ca. 22% less RNA content, (ii) an increased permeability to o-nitrophenyl-beta-d-galactopyranoside (a compound to which untreated cells are impermeable), (iii) almost complete loss of respiratory potential, and (iv) substantial losses of numerous glycolytic enzyme activities in cell extracts prepared from these cells. Intraperiplasmic development of Bdellovibrio bacteriovorus 109J with heat-treated E. coli ML35 as substrate cells appeared normal when observed microscopically, although bdellovibrio attachment and resultant bdelloplast formation were slightly retarded. No significant changes were observed in cell yields or in the ratios and contents of DNA, RNA, or protein between bdellovibrios harvested from untreated cells and those from heat-treated substrate cells after single-developmental-cycle growth on these cells. The average Y(ATP) values for intraperiplasmic growth on untreated and heat-treated substrate cells were 16.0 and 17.9, respectively. It is concluded that intraperiplasmic bdellovibrio growth on gently heat-treated E. coli substrate cells is very similar to growth on untreated substrate cells, even though the former substrate cells are nonviable and substantially impaired in many metabolic activities.

摘要

将大肠杆菌ML35的细胞悬浮液(缓冲液中约每毫升3×10⁹个细胞)进行热处理(55℃处理40分钟),导致细胞活力损失4至4.5个对数级。对其他几种经检测的大肠杆菌菌株也得到了类似结果。当使用用¹⁴C均匀标记的细胞时,经过这种热处理后,260纳米和280纳米吸收物质以及约10%的总细胞放射性被释放到悬浮缓冲液中。与未处理的细胞相比,热处理后的大肠杆菌ML35细胞表现出:(i)除RNA含量约减少22%外,大分子组成无显著变化;(ii)对邻硝基苯基-β-D-吡喃半乳糖苷(一种未处理细胞不能透过的化合物)的通透性增加;(iii)呼吸潜力几乎完全丧失;(iv)从这些细胞制备的细胞提取物中许多糖酵解酶活性大幅丧失。显微镜观察发现,以热处理后的大肠杆菌ML35作为底物细胞时,食菌蛭弧菌109J在周质内的发育看起来正常,尽管蛭弧菌的附着以及由此产生的蛭质体形成略有延迟。在以这些细胞为底物进行单循环生长后,从未处理细胞收获的蛭弧菌与从热处理底物细胞收获的蛭弧菌相比,细胞产量、DNA、RNA或蛋白质的比例和含量均未观察到显著变化。在未处理和热处理的底物细胞上进行周质内生长时,平均Y(ATP)值分别为16.0和17.9。得出的结论是,即使前者底物细胞无活力且许多代谢活动严重受损,但食菌蛭弧菌在轻度热处理的大肠杆菌底物细胞上周质内的生长与在未处理底物细胞上的生长非常相似。

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