源自酿酒酵母菌株W303-1a的设计缺失菌株和原养型菌株。
Designer deletion and prototrophic strains derived from Saccharomyces cerevisiae strain W303-1a.
作者信息
Replogle K, Hovland L, Rivier D H
机构信息
Department of Cell and Structural Biology, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA.
出版信息
Yeast. 1999 Aug;15(11):1141-9. doi: 10.1002/(SICI)1097-0061(199908)15:11<1141::AID-YEA439>3.0.CO;2-P.
We report the construction of Saccharomyces cerevisiae strains isogenic to W303-1a that are designed to allow efficient genetic analysis. To facilitate the generation of null alleles of target genes by PCR-mediated gene disruption, we constructed designer deletion alleles of the ARG4, TRP1 and URA3 genes. In addition, a single pair of oligonucleotide primers were designed that can be used to amplify any of several marker genes for use in PCR-mediated gene disruption. A new version of the 'reusable' hisG-URA3-hisG cassette was constructed for use in PCR-mediated gene disruption. Finally, to facilitate the formation of isogenic diploids by selection, we constructed strains that contain combinations of wild-type alleles of ADE2, HIS3, LEU2, TRP1 and URA3.
我们报告了构建与W303-1a同基因的酿酒酵母菌株，这些菌株旨在便于进行高效的遗传分析。为了通过PCR介导的基因破坏促进靶基因无效等位基因的产生，我们构建了ARG4、TRP1和URA3基因的设计缺失等位基因。此外，设计了一对寡核苷酸引物，可用于扩增几种标记基因中的任何一种，用于PCR介导的基因破坏。构建了一个新版本的“可重复使用”hisG-URA3-hisG盒，用于PCR介导的基因破坏。最后，为了通过选择促进同基因二倍体的形成，我们构建了包含ADE2、HIS3、LEU2、TRP1和URA3野生型等位基因组合的菌株。
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