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酵母絮凝基因FLO1的分子克隆与分析

Molecular cloning and analysis of the yeast flocculation gene FLO1.

作者信息

Watari J, Takata Y, Ogawa M, Sahara H, Koshino S, Onnela M L, Airaksinen U, Jaatinen R, Penttilä M, Keränen S

机构信息

Brewing Research Laboratories, Sapporo Breweries Ltd., Shizuoka, Japan.

出版信息

Yeast. 1994 Feb;10(2):211-25. doi: 10.1002/yea.320100208.

Abstract

The DNA sequence of the flocculation gene FLO1 of Saccharomyces cerevisiae, which is located on chromosome I (Watari et al., 1989) was determined. The sequence contains a large open reading frame (ORF) of 2586 bp and codes for a protein of 862 amino acids. However, further study (genomic Southern and polymerase chain reaction analyses) indicated that the gene we cloned was not the intact FLO1 gene but a form with an approximately 2 kb deletion in the ORF region. The intact FLO1 gene was then cloned and its nucleotide sequence determined. The sequence revealed that the ORF of the intact gene is composed of 4611 bp which code for a protein of 1537 amino acids. A remarkable feature of the putative Flo1 protein is that it contains four families of repeated sequences composed of 18, 2, 3 and 3 repeats and that it has a large number of serines and threonines. In the deleted FLO1 form, a large part of these repeated sequences was missing. The N- and C-terminal regions are hydrophobic and both contain a potential membrane-spanning region, suggesting that the Flo1 protein is an integral membrane protein and a cell wall component.

摘要

酿酒酵母絮凝基因FLO1位于第一条染色体上(Watari等人,1989),其DNA序列已被测定。该序列包含一个2586 bp的大开放阅读框(ORF),编码一个由862个氨基酸组成的蛋白质。然而,进一步研究(基因组Southern杂交和聚合酶链反应分析)表明,我们克隆的基因不是完整的FLO1基因,而是一个在ORF区域有大约2 kb缺失的形式。随后克隆了完整的FLO1基因并测定了其核苷酸序列。该序列显示,完整基因的ORF由4611 bp组成,编码一个由1537个氨基酸组成的蛋白质。推测的Flo1蛋白的一个显著特征是它包含四个由18、2、3和3个重复序列组成的重复序列家族,并且它有大量的丝氨酸和苏氨酸。在缺失的FLO1形式中,这些重复序列的很大一部分缺失了。N端和C端区域是疏水的,都包含一个潜在的跨膜区域,这表明Flo1蛋白是一种整合膜蛋白和细胞壁成分。

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