Fournier P, Guyaneux L, Chasles M, Gaillardin C
Lab. Génétique INRA, Centre de Biotechnologies Agro-Industrielles, Thiverval-Grignon, France.
Yeast. 1991 Jan;7(1):25-36. doi: 10.1002/yea.320070104.
Previous attempts is isolate autonomously replicating sequences (ars) from the dimorphic yeast Yarrowia lipolytica have been unsuccessful. We isolated a Fil- mutant unable to produce hyphae and growing only in a yeast form to facilitate ars isolation. This mutant was transformed with a Y. lipolytica DNA bank and several unstable clones were obtained. Extrachromosomal plasmids were evidenced in yeast, recovered in Escherichia coli and characterized by restriction mapping. They were able to retransform Fil- and Fil+ yeast strains at high frequency and transformants displayed a slightly unstable phenotype. The detailed analysis of the plasmids showed that only two different ars sequences had been isolated, each of them corresponding to a unique sequence in the Y. lipolytica genome. We concluded that functional ars sequences that can be cloned on plasmids are rare in this yeast.
之前从二型酵母解脂耶氏酵母中分离自主复制序列（ars）的尝试均未成功。我们分离出了一个Fil-突变体，它无法产生菌丝，仅以酵母形式生长，以利于ars的分离。用解脂耶氏酵母DNA文库转化该突变体，获得了几个不稳定的克隆。在酵母中证实了存在染色体外质粒，这些质粒在大肠杆菌中得以回收，并通过限制性酶切图谱进行了表征。它们能够高频地重新转化Fil-和Fil+酵母菌株，且转化子表现出稍微不稳定的表型。对这些质粒的详细分析表明，仅分离出了两种不同的ars序列，它们各自对应于解脂耶氏酵母基因组中的一个独特序列。我们得出结论，在这种酵母中，能够克隆到质粒上的功能性ars序列很少见。
