大肠杆菌K12中的基因转化
Genetic transformation in Escherichia coli K12.
作者信息
Cosloy S D, Oishi M
出版信息
Proc Natl Acad Sci U S A. 1973 Jan;70(1):84-7. doi: 10.1073/pnas.70.1.84.
Abstract
An auxotrophic strain of E. coli K12 treated with CaCl(2) was transformed for several markers at a frequency of up to 10(-6) per recipient cell by a DNA preparation isolated from a prototrophic strain. The transforming activity of the DNA preparation was eliminated by treatment with DNase, heat, or sonication, whereas RNase or Pronase treatment had little effect. Two closely linked genetic markers (leu and ara) showed a high degree of cotransformation linkage when high molecular weight DNA was used, but the linkage was almost completely eliminated when sheared, smaller molecular weight DNA was used. There is genetic evidence that the transformation is a result of the replacement of the preexisting genetic marker on the chromosome by that of the donor DNA.
摘要
用氯化钙处理的大肠杆菌K12营养缺陷型菌株，通过从原养型菌株分离的DNA制剂，以每受体细胞高达10^(-6)的频率转化了几个标记。DNA制剂的转化活性经DNA酶、加热或超声处理后被消除，而RNA酶或链霉蛋白酶处理影响很小。当使用高分子量DNA时，两个紧密连锁的遗传标记（亮氨酸和阿拉伯糖）显示出高度的共转化连锁，但当使用剪切后的较小分子量DNA时，这种连锁几乎完全消除。有遗传学证据表明，转化是由于供体DNA的遗传标记取代了染色体上预先存在的遗传标记所致。
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