Kempken F, Kück U
Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Germany.
Mol Gen Genet. 2000 Mar;263(2):302-8. doi: 10.1007/s004380051172.
Restless is an endogenous hAT transposon found in the cyclosporin-producing fungus Tolypocladium inflatum. This element is present in about 15 copies in a particular strain (ATCC34921) which was used for successful gene tagging. We have isolated a T. inflatum mutant with a defect in nitrogen metabolism. This mutant carries a copy of the Restless element in a gene encoding a C6 zinc-finger protein. The deduced amino acid sequence of the gene product shows a significant similarity to the NIT4 protein of Neurospora crassa, which is a regulator of nitrogen metabolism. The wild-type T. inflatum gene was shown to complement a nit-4 mutant of N. crassa. From these data, we conclude that the T. inflatum gene also encodes a regulator of nitrogen metabolism, which was named tnir1 (Tolypocladium nitrogen regulator 1). To the best of our knowledge, this is the first fungal gene to be identified by transposon-directed gene tagging. A general method for gene tagging using an endogenous fungal transposon is presented.
“不安”是一种内源性hAT转座子，存在于产生环孢菌素的真菌多孢霉中。在用于成功进行基因标签的特定菌株（ATCC34921）中，该元件约有15个拷贝。我们分离出了一株氮代谢存在缺陷的多孢霉突变体。该突变体在编码C6锌指蛋白的基因中携带一个“不安”元件拷贝。基因产物推导的氨基酸序列与粗糙脉孢菌的NIT4蛋白有显著相似性，NIT4蛋白是氮代谢的调节因子。野生型多孢霉基因被证明可互补粗糙脉孢菌的nit-4突变体。从这些数据中，我们得出结论，多孢霉基因也编码一种氮代谢调节因子，命名为tnir1（多孢霉氮调节因子1）。据我们所知，这是第一个通过转座子定向基因标签鉴定的真菌基因。本文介绍了一种使用内源性真菌转座子进行基因标签的通用方法。
