Yagi S, Yagi-Tanaka K, Yoshioka J, Suzuki M
Corporate Research and Development Laboratory, Tonen corporation, Saitama, Japan.
Curr Genet. 1993 Jul-Aug;24(1-2):12-20. doi: 10.1007/BF00324659.
We have constructed a series of promoter or upstream activating sequence (UAS)-probe plasmids carrying the Tn5-derived neomycin resistance gene whose seven additional ATG codons in the 5'-untranslated region were completely or partially removed. When the deleted version of the neo sequence retaining only one additional ATG (NeoD) was expressed under the control of a TDH3 promoter whose UAS was deleted, the transformed cells were unable to grow at a low concentration of the antibiotic G418. In contrast with this, yeast cells expressing the NeoC sequence and having no additional ATG exhibited a high level of G418-resistance. Moreover, the UAS-probe system using NeoD has been successfully applied for the identification of several E. coli DNA sequences that clearly function as UASs in yeast cells. Two of these prokaryotic sequences with UAS activity were identified as a part of the coding region of the tgt and the hydG gene, respectively.
我们构建了一系列携带源自Tn5的新霉素抗性基因的启动子或上游激活序列（UAS）探针质粒，该基因5'-非翻译区的七个额外ATG密码子已被完全或部分去除。当仅保留一个额外ATG的neo序列缺失版本（NeoD）在UAS缺失的TDH3启动子控制下表达时，转化细胞在低浓度抗生素G418下无法生长。与此相反，表达NeoC序列且无额外ATG的酵母细胞表现出高水平的G418抗性。此外，使用NeoD的UAS探针系统已成功应用于鉴定几个在酵母细胞中明显作为UAS起作用的大肠杆菌DNA序列。这两个具有UAS活性的原核序列分别被鉴定为tgt和hydG基因编码区的一部分。
