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The REV1 gene of Saccharomyces cerevisiae: isolation sequence and functional analysis.

机译:酿酒酵母的REV1基因:分离序列和功能分析。

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摘要

The REV1 gene of Saccharomyces cerevisiae is required for normal induction of mutations by physical and chemical agents. We have determined the sequence of a 3,485-base-pair segment of DNA that complements the rev1-1 mutant. Gene disruption was used to confirm that this DNA contained the REV1 gene. The sequenced segment contains a single long open reading frame, which can encode a polypeptide of 985 amino acid residues. The REV1 transcript is 3.1 kilobase pairs in length. Frameshift mutations introduced into the open reading frame yielded a Rev-phenotype. A base substitution, encoding Gly-193 to Arg-193, was found in this open reading frame in rev1-1. Deletion mutants, lacking segments of the 5' region of REV1, had intermediate mutability relative to REV1 and rev1-1; a complete deletion exhibited lower mutability than rev1-1. REV1 is not an essential gene. An in-frame fusion of the 5' end of the REV1 open reading frame to the lacZ gene produced beta-galactosidase activity constitutively. The predicted REV1 protein is hydrophilic, with a predicted pI of 9.82. No homologies to RAD1, RAD2, RAD3, RAD7, or RAD10 proteins were noted. A 152-residue internal segment displayed 25% identity with UMUC protein.
机译:酿酒酵母的REV1基因是理化试剂正常诱导突变所必需的。我们已经确定了与rev1-1突变体互补的3,485个碱基对的DNA片段的序列。基因破坏被用于确认该DNA包含REV1基因。测序的片段包含单个长的开放阅读框,其可以编码985个氨基酸残基的多肽。 REV1成绩单的长度为3.1千碱基对。引入开放阅读框的移码突变产生了Rev表型。在rev1-1的此开放阅读框中发现了将Gly-193编码为Arg-193的碱基取代。缺失REV1的5'区域片段的缺失突变体,相对于REV1和rev1-1具有中等的变异性。完全删除的变异性低于rev1-1。 REV1不是必需基因。 REV1开放阅读框的5'端与lacZ基因的框内融合构成了β-半乳糖苷酶活性。预测的REV1蛋白具有亲水性,预测的pI为9.82。没有注意到与RAD1,RAD2,RAD3,RAD7或RAD10蛋白的同源性。 152个残基的内部片段显示与UMUC蛋白的同一性为25%。

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