Grimm C, Schaer P, Munz P, Kohli J
Institute of General Microbiology, University of Bern, Switzerland.
Mol Cell Biol. 1991 Jan;11(1):289-98. doi: 10.1128/mcb.11.1.289-298.1991.
The effect of the strong promoter from the alcohol dehydrogenase gene on mitotic and meiotic intragenic recombination has been studied at the ade6 locus of the fission yeast Schizosaccharomyces pombe. A 700-bp fragment containing the functional adh1 promoter was used to replace the weak wild-type promoter of the ade6 gene. Analysis of mRNA showed that strains with this ade6::adh1 fusion construct had strongly elevated ade6-specific mRNA levels during vegetative growth as well as in meiosis. These increased levels of mRNA correlated with a 20- to 25-fold stimulation of intragenic recombination in meiosis and a 7-fold increased prototroph formation during vegetative growth. Analysis of flanking marker configurations of prototrophic recombinants indicated that simple conversions as well as conversions associated with crossing over were stimulated in meiosis. The strongest stimulation of recombination was observed when the adh1 promoter was homozygous. Studies with heterologous promoter configurations revealed that the highly transcribed allele was the preferred acceptor of genetic information. The effect of the recombinational hot spot mutation ade6-M26 was also investigated in this system. Its effect was only partly additive to the elevated recombination rate generated by the ade6::adh1 fusion construct.
已在裂殖酵母粟酒裂殖酵母的ade6基因座研究了来自乙醇脱氢酶基因的强启动子对有丝分裂和减数分裂基因内重组的影响。用一个包含功能性adh1启动子的700 bp片段替换ade6基因的弱野生型启动子。mRNA分析表明,带有这种ade6::adh1融合构建体的菌株在营养生长以及减数分裂期间ade6特异性mRNA水平大幅升高。这些增加的mRNA水平与减数分裂中基因内重组的20至25倍刺激以及营养生长期间原养型形成增加7倍相关。对原养型重组体侧翼标记构型的分析表明,减数分裂中简单转换以及与交换相关的转换均受到刺激。当adh1启动子纯合时,观察到对重组的最强刺激。对异源启动子构型的研究表明,高转录等位基因是遗传信息的首选受体。还在该系统中研究了重组热点突变ade6-M26的作用。其作用仅部分叠加于ade6::adh1融合构建体产生的升高的重组率。