Development of additional selectable markers for the halophilic Archaeon Haloferax volcanii based on the leuB and trpA genes
Development of additional selectable markers for the halophilic Archaeon Haloferax volcanii based on the leuB and trpA genes
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DOI：
10.1128/aem.70.2.943-953.2004
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发表时间：
2004-02-01
影响因子：
4.4
通讯作者：
Lloyd, RG
中科院分区：
文献类型：
--
作者：
Allers, T;Ngo, HP;Lloyd, RG
由于大多数古生菌是极端的，很难培养，我们目前对它们的生物学知识主要局限于比较基因组学和生物化学。沃氏嗜盐菌作为古细菌遗传学的模式生物有很大的潜力，但到目前为止，这种生物缺乏各种各样的选择标记。我们在这里描述H的分离。volcanii leuB和trpA基因分别编码3-异丙基苹果酸脱氢酶和色氨酸合成酶，并开发这些基因作为阳性选择系统。DeltaleuB和DeltatrpA突变体构建在各种遗传背景，并被证明是亮氨酸和色氨酸营养缺陷型，分别。我们构建了整合型和复制型质粒，在组成型启动子的控制下携带leuB或trpA基因。利用这些选择标记对H. volcanii描述。
Since most archaea are extremophilic and difficult to cultivate, our current knowledge of their biology is confined largely to comparative genomics and biochemistry. Haloferax volcanii offers great promise as a model organism for archaeal genetics, but until now there has been a lack of a wide variety of selectable markers for this organism. We describe here isolation of H. volcanii leuB and trpA genes encoding 3-isopropylmalate dehydrogenase and tryptophan synthase, respectively, and development of these genes as a positive selection system. DeltaleuB and DeltatrpA mutants were constructed in a variety of genetic backgrounds and were shown to be auxotrophic for leucine and tryptophan, respectively. We constructed both integrative and replicative plasmids carrying the leuB or trpA gene under control of a constitutive promoter. The use of these selectable markers in deletion of the lhr gene of H. volcanii is described.