RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama, Japan.
Anal Biochem. 2011 Jul 15;414(2):306-8. doi: 10.1016/j.ab.2011.03.034. Epub 2011 Mar 29.
The expression of foreign proteins in the fission yeast, Schizosaccharomyces pombe, is achieved by introducing an expression vector along with a transducing vector containing an autonomously replicating sequence. We created the expression vector pCMV-Leu2, carrying the LEU2 gene, which complements S. pombeleu1-32, and the transducing vector pUCA-Neo, containing a neomycin-resistance gene. Transformants were screened on leucine-deficient solid medium, followed by rescreening on G418-containing medium. Most of the surviving clones in the initial auxotrophic screening were found to be G418 resistant. The utilization of the pCMV-Leu2 and pUCA-Neo plasmid combination may facilitate rapid screening of S. pombe transformants.
裂殖酵母（Schizosaccharomyces pombe）中外源蛋白的表达是通过引入表达载体和携带自主复制序列的转导载体来实现的。我们构建了表达载体 pCMV-Leu2，携带 LEU2 基因，可补充 S. pombe leu1-32 缺陷；构建了转导载体 pUCA-Neo，携带新霉素抗性基因。通过在缺乏亮氨酸的固体培养基上筛选转化体，然后在含有 G418 的培养基上进行再筛选。在最初的营养缺陷型筛选中，大多数存活的克隆被发现对 G418 有抗性。pCMV-Leu2 和 pUCA-Neo 质粒组合的利用可能有助于快速筛选裂殖酵母转化体。
