Isolation and curing of the Klebsiella pneumoniae large indigenous plasmid using sodium dodecyl sulphate
Isolation and curing of the Klebsiella pneumoniae large indigenous plasmid using sodium dodecyl sulphate
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DOI：
10.1016/s0923-2508(00)00140-6
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发表时间：
2000-04-01
影响因子：
2.6
通讯作者：
Platt, DJ
中科院分区：
文献类型：
--
作者：
El-Mansi, M;Anderson, KJ;Platt, DJ
肺炎克雷伯氏菌（NCTC，CL 687/80）产生并进而分泌谷氨酸的能力等同于存在表观分子量在96 +/- 2 kbp区域内的大型固有质粒（n = 6）。与丝裂霉素C、新生霉素和溴化乙锭（固化剂）不同，十二烷基硫酸钠（SDS）的使用被证明在以相对高的频率（6.25 × 10（-4））固化质粒方面非常有效。此外，在治愈的菌株中不存在异柠檬酸脱氢酶（ICDH）活性强烈表明，与所有已知的ICDH形成鲜明对比的是，在这种生物体中编码ICDH的结构基因是质粒编码的。此外，基于SDS的方案报道了K.肺炎固有质粒已被证明在包括假单胞菌和棒状杆菌的其它生物体中以及在大肠杆菌的重组菌株中是成功的。(C)2000年，Elsevier SAS科学与医学版。
The ability of Klebsiella pneumoniae (NCTC, CL687/80) to produce and, in turn, excrete glutamate has been equated with the presence of a large indigenous plasmid with an apparent molecular mass in the region of 96 +/- 2 kbp (n = 6). Unlike mitomycin C, novobiocine and ethidium bromide (curing agents), the use of sodium dodecyl sulphate (SDS) proved very effective in curing the plasmid with a relatively high frequency (6.25 x 10(-4)). Furthermore, the absence of isocitrate dehydrogenase (ICDH) activity in the cured strain strongly suggests that the structural gene encoding ICDH in this organism, in sharp contrast to all known ICDHs, is plasmid-encoded. Moreover, the SDS-based protocol reported for the isolation of the K. pneumoniae indigenous plasmid has proven successful with other organisms including Pseudomonad and Corynebacteria, as well as in recombinant strains of Escherichia coli. (C) 2000 Editions scientifiques et medicales Elsevier SAS.