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谷氨酸棒杆菌ATCC 13869中ftsZ表达产生的形态改变。

Altered morphology produced by ftsZ expression in Corynebacterium glutamicum ATCC 13869.

作者信息

Ramos Angelina, Letek Michal, Campelo Ana Belén, Vaquera José, Mateos Luis M, Gil José A

机构信息

Departamento de Ecología, Genética y Microbiología, Área de Microbiología, Facultad de Biología, Universidad de León, 24071 León, Spain.

Departamento de Biología Celular, Facultad de Biología, Universidad de León, 24071 León, Spain.

出版信息

Microbiology (Reading). 2005 Aug;151(Pt 8):2563-2572. doi: 10.1099/mic.0.28036-0.

Abstract

Corynebacterium glutamicum is a Gram-positive bacterium that lacks the cell division FtsA protein and actin-like MreB proteins responsible for determining cylindrical cell shape. When the cell division ftsZ gene from C. glutamicum (ftsZ(Cg)) was cloned in different multicopy plasmids, the resulting constructions could not be introduced into C. glutamicum; it was assumed that elevated levels of FtsZ(Cg) result in lethality. The presence of a truncated ftsZ(Cg) and a complete ftsZ(Cg) under the control of Plac led to a fourfold reduction in the intracellular levels of FtsZ, generating aberrant cells displaying buds, branches and knots, but no filaments. A 20-fold reduction of the FtsZ level by transformation with a plasmid carrying the Escherichia coli lacI gene dramatically reduced the growth rate of C. glutamicum, and the cells were larger and club-shaped. Immunofluorescence microscopy of FtsZ(Cg) or visualization of FtsZ(Cg)-GFP in C. glutamicum revealed that most cells showed one fluorescent band, most likely a ring, at the mid-cell, and some cells showed two fluorescent bands (septa of future daughter cells). When FtsZ(Cg)-GFP was expressed from Plac, FtsZ rings at mid-cell, or spirals, were also clearly visible in the aberrant cells; however, this morphology was not entirely due to GFP but also to the reduced levels of FtsZ expressed from Plac. Localization of FtsZ at the septum is not negatively regulated by the nucleoid, and therefore the well-known occlusion mechanism seems not to operate in C. glutamicum.

摘要

谷氨酸棒杆菌是一种革兰氏阳性细菌,它缺乏负责确定圆柱形细胞形状的细胞分裂FtsA蛋白和肌动蛋白样MreB蛋白。当将来自谷氨酸棒杆菌的细胞分裂ftsZ基因(ftsZ(Cg))克隆到不同的多拷贝质粒中时,所得构建体无法导入谷氨酸棒杆菌;据推测,FtsZ(Cg)水平升高会导致致死性。在Plac的控制下,截短的ftsZ(Cg)和完整的ftsZ(Cg)的存在导致FtsZ细胞内水平降低四倍,产生显示出芽、分支和结节但无丝状体的异常细胞。通过用携带大肠杆菌lacI基因的质粒转化使FtsZ水平降低20倍,显著降低了谷氨酸棒杆菌的生长速率,并且细胞更大且呈棒状。对谷氨酸棒杆菌中FtsZ(Cg)的免疫荧光显微镜观察或FtsZ(Cg)-GFP的可视化显示,大多数细胞在细胞中部显示一条荧光带,很可能是一个环,一些细胞显示两条荧光带(未来子细胞的隔膜)。当从Plac表达FtsZ(Cg)-GFP时,在异常细胞中也清晰可见细胞中部的FtsZ环或螺旋;然而,这种形态并不完全归因于GFP,还归因于从Plac表达的FtsZ水平降低。FtsZ在隔膜处的定位不受类核的负调控,因此著名的阻碍机制似乎在谷氨酸棒杆菌中不起作用。

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