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UV Sensitivity and Mutagenesis of the Extremely Thermophilic Eubacterium Thermus thermophilus HB27

机译:极高温嗜热嗜热嗜热杆菌HB27的紫外线敏感性和诱变作用

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References(21) Cited-By(2) Thermus thermophilus is an aerobic, extremely thermophilic eubacterium that grows optimally at 70-75°C. Phylogenetically there is a close relationship between T. thermophilus and Deinococcus radiodurans, an extraordinary radiation-resistant bacterium. In contrast to the D. radiodurans, DNA repair and mutagenesis in T. thermophilus have not been investigated intensively. DNA repair and related mutagenesis at high temperature are particularly interesting because the frequency of DNA damage, such as deamination, depurination, and single strand breaks, is expected to be greatly increased. We disrupted the uvrA gene for subunit A of excinuclease ABC and uvdE gene for probable UV endonuclease by inserting the thermostable kanamycin-resistant gene (HTK) or orotate phosphoribosyltransferase (pyrE) gene. A uvrA mutant showed moderate sensitivity to UV-irradiation, while a uvdE mutant was not UV sensitive. A uvdE uvrA double mutant was highly sensitive to UV-irradiation compared with a uvrA mutant, indicating that UV endonuclease, a uvdE gene product, plays an important role in repair of UV-induced DNA damages. On the other hand, no obvious UV-induced mutations were observed when it was assayed by His+ reversion. It has been reported that T. thermophilus HB27 does not have an SOS response system, because no genes homologous to umuD, umuC, and lexA were found. In the absence of error-prone translesion DNA polymerases in T. thermophilus, mutations would not occur at the site of pyrimidine dimers
机译:参考文献(21)By-By(2)Thermus thermophilus是一种需氧的,高度嗜热的真细菌,在70-75°C最佳生长。从系统发育上讲,嗜热链球菌与耐辐射的奇异球菌之间存在密切的关系。与D. radiodurans相比,嗜热衣原体的DNA修复和诱变尚未得到深入研究。高温下的DNA修复和相关诱变特别有趣,因为DNA损伤(如脱氨,脱嘌呤和单链断裂)的发生频率有望大大提高。通过插入热稳定的卡那霉素抗性基因(HTK)或乳清酸磷酸核糖基转移酶(pyrE)基因,我们破坏了核酸外切酶ABC A亚基的uvrA基因和可能的紫外线核酸内切酶的uvdE基因。 uvrA突变体对紫外线照射显示中等灵敏度,而uvdE突变体对紫外线不敏感。 uvdE uvrA双突变体与uvrA突变体相比对UV辐射高度敏感,这表明uvdE基因产物UV内切核酸酶在修复UV诱导的DNA损伤中起重要作用。另一方面,当用His +还原法检测时,没有观察到明显的紫外线诱导的突变。据报道,由于没有发现与umuD,umuC和lexA同源的基因,嗜热链球菌HB27没有SOS应答系统。在嗜热链球菌中不存在容易出错的易转移DNA聚合酶的情况下,在嘧啶二聚体的位点不会发生突变

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