Spores harboring an ACC1 deletion derived from a diploid Saccharomyces cerevisiae strain, in which one copy of the entire ACC1 gene is replaced with a LEU2 cassette, fail to grow. A chimeric gene consisting of the yeast GAL10 promoter, yeast ACC1 leader, wheat cytosolic acetyl-CoA carboxylase (ACCase) cDNA, and yeast ACC1 3' tail was used to complement a yeast ACC1 mutation. The complementation demonstrates that active wheat ACCase can be produced in yeast. At low concentrations of galactose, the activity of the "wheat gene" driven by the GAL10 promoter is low and ACCase becomes limiting for growth, a condition expected to enhance transgenic yeast sensitivity to wheat ACCase-specific inhibitors. An aryloxyphenoxypropionate and two cyclohexanediones do not inhibit growth of haploid yeast strains containing the yeast ACC1 gene, but one cyclohexanedione inhibits growth of the gene-replacement strains at concentrations below 0.2 mM. In vitro, the activity of wheat cytosolic ACCase produced by the gene-replacement yeast strain is inhibited by haloxyfop and cethoxydim at concentrations above 0.02 mM. The activity of yeast ACCase is less affected. The wheat plastid ACCase in wheat germ extract is inhibited by all three herbicides at concentrations below 0.02 mM. Yeast gene-replacement strains will provide a convenient system for the study of plant ACCases.

译文

:带有源自二倍体酿酒酵母菌株的ACC1缺失的孢子无法生长,其中整个ACC1基因的一个拷贝被LEU2盒代替。由酵母GAL10启动子,酵母ACC1前导序列,小麦胞质乙酰辅酶A羧化酶(ACCase)cDNA和酵母ACC1 3'尾部组成的嵌合基因用于补充酵母ACC1突变。互补表明,可以在酵母中产生活性小麦ACCase。在低浓度的半乳糖下,由GAL10启动子驱动的“小麦基因”的活性低,ACCase成为限制生长的条件,该条件有望增强转基因酵母对小麦ACCase特异性抑制剂的敏感性。芳氧基苯氧基丙酸酯和两种环己二酮不抑制含有酵母ACC1基因的单倍体酵母菌株的生长,但是一种环己二酮在低于0.2 mM的浓度下抑制基因置换菌株的生长。在体外,由基因替代酵母菌株产生的小麦胞质ACCase的活性在0.02 mM以上的浓度下被haloxyfop和cethoxydim抑制。酵母ACCase的活性受到的影响较小。浓度低于0.02 mM的所有三种除草剂均能抑制小麦胚芽提取物中的小麦质体ACCase。酵母基因置换菌株将为植物ACCases的研究提供方便的系统。

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