Transformation of the methylotrophic yeast Hansenula polymorpha
Transformation of the methylotrophic yeast Hansenula polymorpha
复制标题
甲基营养酵母汉逊酵母的转化
DOI：
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发表时间：
1986
期刊：
影响因子：
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通讯作者：
P. Sudbery
中科院分区：
文献类型：
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作者：
M. Gleeson;G. S. Ortori;P. Sudbery
摘要：分离到一株β-异丙基苹果酸脱氢酶缺陷突变体。用含有酿酒酵母LEU2基因的穿梭载体YEp13分别用原生质体法和LiCl处理对其进行转化。以30-60 μg (DNA)-1的频率回收Leu+转化子。通过稳定性测试、杂交实验以及通过转化大肠杆菌从转化细胞中回收YEp13，证明转化子含有自主复制的质粒。经BamHI消化线性化后的YEp13也能有效转化；线性化的质粒可以在转化体中自动循环和复制。
Summary: A mutant of Hansenula polymorpha defective in β-isopropylmalate dehydrogenase was isolated. It was transformed with YEp13, a shuttle vector containing the Saccharomyces cerevisiae LEU2 gene, either using the protoplasting method or by treatment with LiCl. Leu+ transformants were recovered at a frequency of 30-60 μg DNA)-1. The transformants were shown to contain an autonomously replicating plasmid by stability tests, hybridization experiments and by the recovery of YEp13 from transformed cells via transformation of Escherichia coli. YEp13 linearized by BamHI digestion was also effective in transformation; the linearized plasmid was shown to recircularize and replicate autonomously in the transformants.