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转座子诱变揭示了影响盐胁迫耐受性和低温生长的新基因座。

Transposon mutagenesis reveals novel loci affecting tolerance to salt stress and growth at low temperature.

作者信息

de Jesus Ferreira M C, Bao X, Laizé V, Hohmann S

机构信息

Department of Cell and Molecular Biology/Microbiology, Göteborg University, Sweden.

出版信息

Curr Genet. 2001 Aug;40(1):27-39. doi: 10.1007/s002940100237.

Abstract

Using transposon mutagenesis in the haploid Saccharomyces cerevisiae strain W303-1A we have identified genes required for growth in high salt medium, survival of a hypo-osmotic shock and growth at 15 degrees C. Screening 25,000 transposon insertions revealed a total of 61 insertions that caused salt-sensitivity; and those insertions affected 31 genes. Only 12 of those genes were previously known to be required for salt-tolerance. Among the 61 insertions, three caused general osmo-sensitivity. We identified one single insertion mutant in the already-known gene, FPS1, required for survival of hypo-osmotic shock. A total of 31 insertions caused failure to grow at low temperature. Those identified ten different genes, three of which had previously been reported to affect cold-tolerance. Four genes were identified in both the salt and the cold-sensitivity screen. We found several unusual insertion mutations: (1) insertions in or close to essential genes, (2) insertion in an intergenic region and (3) insertions causing stress-sensitivity in W303-1A, while the deletion mutant in BY4741 did not show such a phenotype. Surprisingly, our mutant set and that reported in the large-scale transposon insertion project (TRIPLES, http://ygacmed.yale.edu/triples/triples.htm) only marginally overlap. We discuss some of the features of transposon mutagenesis in light of the availability of the complete set of yeast deletion mutants and we discuss the possible roles of the genes we identified.

摘要

利用单倍体酿酒酵母菌株W303-1A中的转座子诱变技术,我们鉴定出了在高盐培养基中生长、低渗休克存活以及在15摄氏度下生长所需的基因。对25,000个转座子插入进行筛选,共发现61个导致盐敏感性的插入;这些插入影响了31个基因。其中只有12个基因先前已知是耐盐所必需的。在这61个插入中,有3个导致普遍的渗透敏感性。我们在已知的低渗休克存活所需基因FPS1中鉴定出一个单插入突变体。共有31个插入导致在低温下无法生长。这些鉴定出了10个不同的基因,其中3个先前已报道影响耐寒性。在盐敏感性和冷敏感性筛选中都鉴定出了4个基因。我们发现了几个不寻常的插入突变:(1) 在必需基因内或其附近的插入,(2) 在基因间区域的插入,以及(3) 在W303-1A中导致应激敏感性的插入,而BY4741中的缺失突变体未表现出这种表型。令人惊讶的是,我们的突变体集合与大规模转座子插入项目(TRIPLES,http://ygacmed.yale.edu/triples/triples.htm)中报道的突变体集合仅有少量重叠。我们根据酵母缺失突变体全套的可得性讨论了转座子诱变的一些特征,并讨论了我们鉴定出的基因的可能作用。

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