REPLICATIVE TRANSFORMATION OF THE FILAMENTOUS FUNGUS ASHBYA-GOSSYPII WITH PLASMIDS CONTAINING SACCHAROMYCES-CEREVISIAE ARS ELEMENTS
REPLICATIVE TRANSFORMATION OF THE FILAMENTOUS FUNGUS ASHBYA-GOSSYPII WITH PLASMIDS CONTAINING SACCHAROMYCES-CEREVISIAE ARS ELEMENTS
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DOI：
10.1016/0378-1119(91)90593-z
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发表时间：
1991-12-20
期刊：
影响因子：
3.5
通讯作者：
PHILIPPSEN, P
中科院分区：
文献类型：
--
作者：
WRIGHT, MC;PHILIPPSEN, P
我们开发了一种丝状子囊菌真菌棉丝Ashbya gossypii转化系统。利用含有大肠杆菌卡那霉素耐药基因的质粒作为选择性标记，并从酿酒酵母（Saccharomyces cerevisiae）中自主复制序列（ARS），将菌丝原生质体转化为耐药菌株。每mu-g质粒DNA的转化频率高达63个转化子。在非选择条件下，变形子是不稳定的。对常规和脉冲场凝胶电泳分离的DNA进行南方分析表明，转化DNA以自主复制质粒的形式存在。没有检测到质粒与染色体DNA的整合。我们的结论是，葡萄球菌ARS元件在棉蚜中是功能性的，因为缺乏这些元件的载体不能产生转化子。
We have developed a transformation system for the filamentous ascomycete fungus Ashbya gossypii. Mycelial protoplasts were transformed to geneticin-resistance with plasmids containing the Escherichia coli kanamycin-resistance gene as a selectable marker and autonomously replicating sequences (ARS) from Saccharomyces cerevisiae (ARS1, 2-mu-ARS). Transformation frequencies of up to 63 transformants per mu-g of plasmid DNA were obtained. The transformants were unstable under nonselective conditions. Southern analysis of DNA separated by conventional and pulsed-field-gel electrophoresis showed that the transforming DNA was present as autonomously replicating plasmid. Plasmid integration into chromosomal DNA was not detected. We concluded that the S. cerevisiae ARS elements are functional in A. gossypii, since vectors lacking such elements did not yield transformants.