The SSK2 MAPKKK of Candida albicans is required for oxidant adaptation in vitro.
The SSK2 MAPKKK of Candida albicans is required for oxidant adaptation in vitro.
复制标题
白色念珠菌的 SSK2 MAPKKK 是体外氧化剂适应所必需的。
DOI：
10.1111/j.1567-1364.2007.00329.x
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发表时间：
2008
影响因子：
3.2
通讯作者：
Calderone,Richard
中科院分区：
文献类型：
--
作者：
Walia,Aditi;Calderone,Richard
白色念珠菌的 Ssk2p (MAPKKK) 被删除，并根据表型研究分配功能。首先使用 UAU1 破坏方法尝试删除 SSK2。所有缺乏一个SSK2拷贝的转化体似乎都是三倍体，这表明SSK2对于该生物体是必需的。为了验证这一观察结果，构建了一种菌株，其中使用 SAT1flipper 破坏方法删除了一个等位基因。然后将第二个等位基因置于开/关四环素调节（TetR）启动子的控制之下。通过逆转录酶-PCR测量SSK2的转录，尽管启动子有些渗漏，但在多西环素存在下，anssk2/TetR-SSK2转化子(AT2)中的转录显着减少。分别使用SAT1flipper和TetR启动子方法构建的菌株AT1和AT2在不同生长培养基中进行表型研究以确定Ssk2p在形态发生中的作用。还在 1.5 M 氯化钠和各种类型氧化剂存在的情况下，在开/关条件下对突变体进行了比较。菌株 AT2 在没有强力霉素的情况下表现出对 1.5 M NaCl 的抗性，但被 8 mM 过氧化氢抑制。
The Ssk2p (MAPKKK) ofCandida albicanswas deleted and functions assigned based on phenotyping studies.SSK2deletion was first attempted using the UAU1 disruption method. All transformants lacking one copy ofSSK2appeared to be triploids, suggesting that theSSK2is essential for the organism. To verify this observation, a strain was constructed in which one allele was deleted using theSAT1flipper disruption method. The second allele was then placed under control of the on/off tetracycline-regulatable (TetR) promoter. The transcription ofSSK2was measured by reverse transcriptase-PCR and although the promoter was somewhat leaky, transcript was significantly reduced in anssk2/TetR-SSK2transformant (AT2) in the presence of doxycycline. Strains AT1 and AT2 constructed using theSAT1flipper and TetR promoter method, respectively, were studied phenotypically in different growth media to determine the role of Ssk2p in morphogenesis. The mutants were also compared under on/off conditions in the presence of 1.5 M NaCl and various types of oxidants. Strain AT2 demonstrated resistance to 1.5 M NaCl in the absence of doxycycline but was inhibited by 8 mM hydrogen peroxide.