Das S, Kellermann E, Hollenberg C P
J Bacteriol. 1984 Jun;158(3):1165-7. doi: 10.1128/jb.158.3.1165-1167.1984.
For the transformation of the yeast species Kluyveromyces fragilis, we have constructed a vector containing a bacterial kanamycin resistance (Kmr) gene, the TRP1 gene of Saccharomyces cerevisiae, and an autonomously replicating sequence of Kluyveromyces lactis called KARS2 . By utilizing the method based on treatment by alkali cations and with the Kmr gene as the selective marker, a wild-type strain of K. fragilis was transformed to resistance against the antibiotic G418 . In the transformed cell the plasmid replicates autonomously. The same plasmid could also be used to transform S. cerevisiae trp1 mutant to Trp+. Thus, KARS2 of K. lactis enables the vector to replicate in K. fragilis, K. lactis, and S. cerevisiae, whereas ARS1 of S. cerevisiae allows autonomous replication only in S. cerevisiae.
为了实现脆壁克鲁维酵母的转化，我们构建了一个载体，该载体包含一个细菌卡那霉素抗性（Kmr）基因、酿酒酵母的TRP1基因以及乳酸克鲁维酵母的一个自主复制序列KARS2。通过利用基于碱金属阳离子处理的方法，并以Kmr基因作为选择标记，脆壁克鲁维酵母的野生型菌株被转化为对抗生素G418具有抗性。在转化细胞中，质粒自主复制。同样的质粒也可用于将酿酒酵母trp1突变体转化为Trp+。因此，乳酸克鲁维酵母的KARS2使该载体能够在脆壁克鲁维酵母、乳酸克鲁维酵母和酿酒酵母中复制，而酿酒酵母的ARS1仅允许在酿酒酵母中自主复制。
