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首页> 外文期刊>Journal of bacteriology >Chimeric plasmids for cloning of deoxyribonucleic acid sequences in Saccharomyces cerevisiae.
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Chimeric plasmids for cloning of deoxyribonucleic acid sequences in Saccharomyces cerevisiae.

机译：用于在酿酒酵母中克隆脱氧核糖核酸序列的嵌合质粒。

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Two sets of plasmids, each carrying a Saccharomyces cerevisiae gene and a portion or all of the yeast 2-micron circle linked to the Escherichia coli plasmid pBR322, have been constructed. One of these sets contains a BamHI fragment of S. cerevisiae deoxyribonucleic acid that includes the yeast his3 gene, whereas the other set contains a BamHI fragment of S. cerevisiae that includes the yeast leu2 gene. All plasmids transform S. cerevisiae and E. coli with a high frequency, possess unique restriction endonuclease sites, and are retrievable from both host organisms. Plasmids carrying the 2.4-megadalton EcoRI fragment of the 2-micron circle transform yeast with 2- to 10-fold greater frequency than those carrying the 1.5-megadalton EcoRI fragment of the 2-micron circle. Restriction endonuclease analysis of plasmics retrieved from S. cerevisiae transformed with plasmics carrying the 2.4-megadalton EcoRI fragment showed that in 13 of 96 cases the original plasmic has acquired an additional copy of the 2-mcron circle. These altered plasmids appear to have arisen by means of an interplasmid recombination event while in S. cerevisiae. A clone bank of S. cerevisiae genes based upon one of these composite plasmids has been constructed. By using this bank and selecting directly in S. cerevisiae, the ura3, tyr1, and met2 genes have been cloned.
机译：已经构建了两组质粒，每组均携带啤酒酵母基因和与大肠杆菌质粒pBR322连接的部分或全部酵母2微米环。这些组中的一组包含啤酒酵母脱氧核糖核酸的BamHI片段，其包括酵母his3基因，而另一组包含啤酒酵母的BamHI片段，其包括酵母leu2基因。所有质粒均以高频率转化酿酒酵母和大肠杆菌，具有独特的限制性核酸内切酶位点，并且可从两种宿主生物中检索到。携带2微米圈的2.4兆达尔顿EcoRI片段的质粒比携带2微米圈的1.5兆达尔顿EcoRI片段的质粒将酵母转化的频率高2至10倍。对从携带有2.4兆达尔顿EcoRI片段的质粒转化的啤酒酵母中检索到的质粒进行限制性核酸内切酶分析，结果表明，在96个病例中，有13例原始质粒获得了额外的2 mcron环。这些改变的质粒似乎是在酿酒酵母中通过质粒间重组事件产生的。已经构建了基于这些复合质粒之一的酿酒酵母基因的克隆库。通过使用该库并直接在酿酒酵母中进行选择，已克隆了ura3，tyr1和met2基因。

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