Lennon E, Minton K W
Department of Pathology, F. E. Hebert School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799.
J Bacteriol. 1990 Jun;172(6):2955-61. doi: 10.1128/jb.172.6.2955-2961.1990.
Deinococcus radiodurans is the most-studied species of a eubacterial family characterized by extreme resistance to DNA damage. We have focused on developing molecular biological techniques to investigate the genetics of this organism. We report construction of lacZ gene fusions by a method involving both in vitro splicing and the natural transformation of D. radiodurans. Numerous fusion strains were identified by expression of beta-galactosidase. Among these fusion strains, several were inducible by exposure to the DNA-damaging agent mitomycin C, and four of the inducible fusion constructs were cloned in Escherichia coli. Hybridization studies indicate that one of the damage-inducible genes contains a sequence reiterated throughout the D. radiodurans chromosome. Survival measurements show that two of the fusion strains have increased sensitivity to mitomycin C, suggesting that the fusions within these strains inactivate repair functions.
耐辐射球菌是真细菌家族中研究最多的物种，其特点是对DNA损伤具有极强的抗性。我们专注于开发分子生物学技术来研究这种生物体的遗传学。我们报告了通过一种涉及体外剪接和耐辐射球菌自然转化的方法构建lacZ基因融合体。通过β-半乳糖苷酶的表达鉴定出了许多融合菌株。在这些融合菌株中，有几种可通过暴露于DNA损伤剂丝裂霉素C诱导，并且其中四个可诱导融合构建体在大肠杆菌中进行了克隆。杂交研究表明，其中一个损伤诱导基因包含在耐辐射球菌染色体中反复出现的序列。存活测量表明，其中两个融合菌株对丝裂霉素C的敏感性增加，这表明这些菌株中的融合使修复功能失活。
