TRANSFORMATION OF INTACT YEAST-CELLS TREATED WITH ALKALI CATIONS
TRANSFORMATION OF INTACT YEAST-CELLS TREATED WITH ALKALI CATIONS
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DOI：
10.1128/jb.153.1.163-168.1983
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发表时间：
1983-01-01
影响因子：
3.2
通讯作者：
KIMURA, A
中科院分区：
文献类型：
--
作者：
ITO, H;FUKUDA, Y;KIMURA, A
用碱金属阳离子处理的完整酵母细胞摄取质粒DNA。Li+、Cs+、Rb+、K+和Na+对感受性有诱导作用。S.用CsCl对酵母D13-1A与质粒YRp 7的融合进行了研究。最适孵育时间为1小时，最适细胞浓度为5 × 104。107个细胞/ml。Cs+的最适浓度为1.0M。转化效率随着质粒DNA浓度的增加而增加。聚乙二醇是绝对必要的。热脉冲和各种多胺或碱性蛋白刺激质粒DNA的摄取。除了环状DNA，线性质粒DNA也采取了Cs+处理的酵母细胞，虽然摄取效率大大降低。对于含有ars 1的质粒，用Cs+或Li+的转化效率与常规原生质体方法的转化效率相当，但对于含有2 μ m复制起点的质粒则不然。
Intact yeast cells treated with alkali cations took up plasmid DNA. Li+, Cs+, Rb+, K+ and Na+ were effective in inducing competence. Conditions for the transformation of S. cerevisiae D13-1A with plasmid YRp7 were studied in detail with CsCl. The optimum incubation time was 1 h, and the optimum cell concentration was 5 .times. 107 cells/ml. The optimum concentration of Cs+ was 1.0 M. Transformation efficiency increased with increasing concentrations of plasmid DNA. Polyethylene glycol was absolutely required. Heat pulse and various polyamines or basic proteins stimulated the uptake of plasmid DNA. Besides circular DNA, linear plasmid DNA was also taken up by Cs+-treated yeast cells, although the uptake efficiency was considerably reduced. The transformation efficiency with Cs+ or Li+ was comparable with that of conventional protoplast methods for a plasmid containing ars1, although not for plasmids containing a 2.mu.m origin replication.