CLONING OF YEAST GENE FOR TRICHODERMIN RESISTANCE AND RIBOSOMAL PROTEIN-L3
CLONING OF YEAST GENE FOR TRICHODERMIN RESISTANCE AND RIBOSOMAL PROTEIN-L3
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DOI：
10.1073/pnas.78.1.238
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
WARNER, JR
中科院分区：
文献类型：
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作者：
FRIED, HM;WARNER, JR
用携带抗霉菌素酵母基因组DNA片段的自主复制重组质粒转化对真核蛋白质合成抑制物霉菌素敏感的酵母细胞。经过对毛霉素抗性细胞的筛选，几个转化子产生了一个含有13.5kb DNA片段的质粒，该DNA片段编码毛霉素抗性基因tcm1和酵母核糖体蛋白中最大的核糖体蛋白L3基因。携带该质粒的细胞对毛霉菌素、相关药物疣菌素A和无关药物山奈素具有抗药性。这种耐药模式与携带tcm1染色体拷贝的菌株所表现出的相似。从转化细胞制备的多核糖体在体外蛋白质合成试验中对毛霉菌素具有抗药性。对13.5kb的DNA片段进行亚克隆，发现Tcm1基因和L3蛋白基因均包含在3.2kb的片段中。酵母中耐药木霉素的基因是核糖体蛋白L3。
Yeast cells sensitive to the eukaryotic protein synthesis inhibitor trichodermin were transformed with autonomously replicating recombinant plasmids carrying DNA fragments of the genome of a trichodermin-resistant yeast strain. After selection for trichodermin-resistant cells, several transformants yielded a plasmid containing a 13.5 kilobase (kb) DNA fragment that encodes the trichodermin resistance gene, tcm1 and the gene for ribosomal protein L3, the largest of the yeast ribosomal proteins. Cells carrying this plasmid are resistant to trichodermin and to the related drug verrucarin A and to the unrelated drug anisomycin. This pattern of resistance is similar to that exhibited by strains carrying a chromosomal copy of tcm1. Polyribosomes prepared from transformed cells are resistant to trichodermin when tested in an in vitro protein synthesis assay. Subcloning of the 13.5 kb DNA fragment revealed that the gene for tcm1 and the gene for protein L3 are contained within a 3.2 kb segment. The gene for trichodermin resistance in yeast specifies ribosomal protein L3.