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酿酒酵母SEC1基因的克隆与测序,该基因位于第四条染色体上。

Cloning and sequencing of the yeast Saccharomyces cerevisiae SEC1 gene localized on chromosome IV.

作者信息

Aalto M K, Ruohonen L, Hosono K, Keränen S

机构信息

VTT, Biotechnical Laboratory, Espoo, Finland.

出版信息

Yeast. 1991 Aug-Sep;7(6):643-50. doi: 10.1002/yea.320070613.

Abstract

The SEC1 gene of yeast Saccharomyces cerevisiae was cloned by complementing the temperature-sensitive mutation of sec1-1 at 37 degrees C, and its nucleotide sequence was determined. SEC1 is a single copy gene and encodes a protein of 724 amino acids and 83,490 daltons with a predicted pI value of 6.11. Hydrophobicity plotting showed no clearly hydrophobic regions suggesting a soluble nature for the protein. Amino acid sequence comparisons revealed no obvious homologies with the proteins in the SWISSPROT databank. Two consensus sequence for the cdc2 encoded protein kinase recognition site were revealed within Sec1p. The codon usage suggests a low expression level for SEC1. The 5' non-translated region contains two TATA-like sequences at -52 and -215 nucleotides from the translation start site. Two potential regulatory sequences for DNA binding proteins were found in the non-coding 5' region: a HAP2/HAP3 consensus recognition sequence at nucleotide-154 and a BAF1 consensus recognition sequence at nucleotide-136. The SEC1 specific probe detected a 2400 nucleotides long transcript, which was in reasonable agreement with the 2172 nucleotides long open reading frame.

摘要

通过在37℃下对酵母酿酒酵母的温度敏感突变体sec1 - 1进行互补克隆了SEC1基因,并测定了其核苷酸序列。SEC1是一个单拷贝基因,编码一种由724个氨基酸组成、分子量为83490道尔顿的蛋白质,预测的pI值为6.11。疏水性绘图显示没有明显的疏水区域,表明该蛋白质具有可溶性。氨基酸序列比较显示与SWISSPROT数据库中的蛋白质没有明显的同源性。在Sec1p中发现了cdc2编码的蛋白激酶识别位点的两个共有序列。密码子使用情况表明SEC1的表达水平较低。5'非翻译区在距翻译起始位点-52和-215个核苷酸处含有两个类似TATA的序列。在非编码5'区域发现了两个潜在的DNA结合蛋白调控序列:核苷酸-154处的HAP2/HAP3共有识别序列和核苷酸-136处的BAF1共有识别序列。SEC1特异性探针检测到一个2400个核苷酸长的转录本,这与2172个核苷酸长的开放阅读框合理一致。

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