Fernández-Herrero L A, Olabarría G, Castón J R, Lasa I, Berenguer J
Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid-Consejo, Spain.
J Bacteriol. 1995 Oct;177(19):5460-6. doi: 10.1128/jb.177.19.5460-5466.1995.
The S-layers of Thermus thermophilus HB27 and T. thermophilus HB8 are composed of protein units of 95 kDa (P95) and 100 kDa (P100), respectively. We have selected S-layer deletion mutants from both strains by complete replacement of the slpA gene. Mutants of the two strains showed similar defects in growth and morphology and overproduced an external cell envelope inside of which cells remained after division. However, the nature of this external layer is strain specific, being easily stained and regular in the HB8 delta slpA derivative and amorphous and poorly stained in the HB27 delta slpA strain. The addition of chromosomic DNA from T. thermophilus HB8 to growing cultures of T. thermophilus HB27 delta slpA led to the selection of a new strain, HB27C8, which expressed a functional S-layer composed of the P100 protein. Conversely, the addition of chromosomic DNA from T. thermophilus HB27 to growing cultures of T. thermophilus HB8 delta slpA allowed the isolation of strain HB8C27, which expressed a functional S-layer composed of the P95 protein. The driving force which selected the transference of the S-layer genes in these experiments was the difference in growth rates, one of the main factors leading to selection in natural environments.
嗜热栖热菌HB27和嗜热栖热菌HB8的S层分别由95 kDa的蛋白质单元(P95)和100 kDa的蛋白质单元(P100)组成。我们通过完全替换slpA基因从这两个菌株中筛选出了S层缺失突变体。这两个菌株的突变体在生长和形态上表现出相似的缺陷,并且过量产生了一种外部细胞包膜,细胞分裂后仍留在其中。然而,这种外层的性质具有菌株特异性,在HB8 δslpA衍生物中易于染色且规则,而在HB27 δslpA菌株中则是无定形的且染色较差。将嗜热栖热菌HB8的染色体DNA添加到嗜热栖热菌HB27 δslpA的生长培养物中,导致筛选出了一个新菌株HB27C8,该菌株表达了由P100蛋白组成的功能性S层。相反,将嗜热栖热菌HB27的染色体DNA添加到嗜热栖热菌HB8 δslpA的生长培养物中,使得分离出了菌株HB8C27,该菌株表达了由P95蛋白组成的功能性S层。在这些实验中选择S层基因转移的驱动力是生长速率的差异,这是自然环境中导致选择的主要因素之一。