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首页> 外文学位 >Isolation and characterization of ELO1, a gene required for fatty acid elongation in Saccharomyces cerevisiae.
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Isolation and characterization of ELO1, a gene required for fatty acid elongation in Saccharomyces cerevisiae.

机译：ELO1的分离和鉴定，ELO1是酿酒酵母中脂肪酸延伸所需的基因。

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摘要

A fatty acid elongation defective mutant was isolated from Saccharomyces cerevisiae by mutagenizing strains that were defective in fatty acid synthase (FAS) activity. Cells of the fatty acid synthase-defective strains can grow when supplemented with tetradecanoic acid (14:0) due to the presence of membrane bound elongation systems that can extend the 14-carbon fatty acid to longer-chain species. After mutagenesis and rescue on medium containing a mixture of 14:0, 16:0 and 18:0, cells were screened for their inability to grow on medium containing only 14:0. From 150,000 colonies, 4 stable isolates were identified, all of which appear to represent the same complementation group. Based on these analyses, the mutant isolates are all believed to carry the same defective gene elo1-1 (designated as elongation defective).; Gas chromatography of lipid extracts from cells of a fas2{dollar}Delta {dollar};elo1-1 mutant (DTY10m2) indicates that it fails to efficiently elongate (12, 13 or 14) carbon fatty acids. A gene disrupted fas 2{dollar}Delta{dollar}::LEU2; elo1{dollar}Delta{dollar}::HIS3 mutant incorporates 14 to 18-carbon fatty acids into membrane lipids, indicating that fatty acid transport is not affected by the mutation. Analysis of the ELO1 mRNA levels indicates that the gene is expressed in cells grown on fatty acid deficient medium. It is rapidly induced in wild type cells that are supplemented with 14:0 and is repressed when cells are supplied with 16 and 18-carbon fatty acids. Molecular cloning and sequence analysis of the ELO1 gene suggests that the encoded protein is a membrane bound polypeptide that contains at least 5 potential membrane spanning regions and a presumptive NADPH binding site. Homology searches of the yeast database indicate the existence of at least 2 other genes that share greater than 50% identity with ELO1. These genes are currently under investigation to determine their roles in fatty acid elongation.; The yeast ELO1 gene represents a novel pathway in the elongation of tetradecanoic acid (14:0) to that of hexadecanoic acid (16:0), distinct from that of fatty acid synthase.
机译：通过诱变在脂肪酸合酶（FAS）活性方面有缺陷的菌株，从酿酒酵母中分离出一种脂肪酸延伸缺陷突变体。脂肪酸合酶缺陷菌株的细胞在补充十四烷酸（14：0）时可以生长，这是因为存在膜结合的延伸系统，该系统可以将14个碳原子的脂肪酸延伸至更长链的物种。在包含14：0、16：0和18：0混合物的培养基上诱变并营救后，筛选细胞无法在仅包含14：0的培养基上生长的能力。从150,000个菌落中，鉴定出4个稳定的分离株，它们似乎都代表相同的互补组。基于这些分析，认为突变株均携带相同的缺陷基因elo1-1（被指定为延伸缺陷）。气相色谱法分析fas2 {delta} {elo1-1; elo1-1突变体（DTY10m2）的细胞中的脂质提取物，它无法有效地延长（12、13或14）个碳脂肪酸。一个基因破坏了fas 2 {dollar} Delta {dollar} :: LEU2； elo1 {dollar} Delta {dollar} :: HIS3突变体将14至18个碳的脂肪酸掺入膜脂质中，表明脂肪酸运输不受突变影响。对ELO1 mRNA水平的分析表明，该基因在脂肪酸缺乏培养基上生长的细胞中表达。它在补充14：0的野生型细胞中迅速诱导，并在向细胞提供16和18碳脂肪酸的情况下被抑制。 ELO1基因的分子克隆和序列分析表明，编码的蛋白是一种膜结合多肽，包含至少5个潜在的跨膜区域和一个推测的NADPH结合位点。酵母数据库的同源性搜索表明存在至少2个与ELO1的同一性大于50％的其他基因。目前正在研究这些基因以确定它们在脂肪酸延伸中的作用。酵母ELO1基因代表十四烷酸（14：0）延伸到十六烷酸（16：0）的新途径，这与脂肪酸合酶的途径不同。

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  • 作者

    Toke, David Alexander.;

  • 作者单位

    Rutgers The State University of New Jersey - New Brunswick.;

  • 授予单位 Rutgers The State University of New Jersey - New Brunswick.;
  • 学科 Biology Molecular.; Biology Genetics.; Biology Cell.
  • 学位 Ph.D.
  • 年度 1996
  • 页码 169 p.
  • 总页数 169
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;遗传学;细胞生物学;
  • 关键词

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