LOCALIZED MUTAGENIS OF ANY SPECIFIC SMALL REGION OF BACTERIAL CHROMOSOME
LOCALIZED MUTAGENIS OF ANY SPECIFIC SMALL REGION OF BACTERIAL CHROMOSOME
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DOI：
10.1073/pnas.68.12.3158
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发表时间：
1971-01-01
影响因子：
11.1
通讯作者：
AMES, BN
中科院分区：
文献类型：
--
作者：
HONG, JS;AMES, BN
本文描述了一种方法，我们称之为局部诱变，用于分离细菌染色体上任何特定小区域（约1%）的温度敏感突变和其他类型的突变。该方法的原理是突变转导DNA而不是细菌DNA。人们可以通过将该区域中的营养缺陷型标记转导为原养型，从而在邻近区域中引入新的突变，来选择将这种突变的DNA引入细菌染色体的任何特定区域。我们已经用这种方法分离出许多不同的温度敏感性突变基因的未知功能的特定区域的染色体。由于该方法非常简单，因此可用于使图谱的任何区域都充满必需基因的突变，或用于各种类型的遗传操作。虽然我们已经用羟胺诱变的噬菌体P22和鼠伤寒沙门氏菌进行了研究，但该方法也适用于其他诱变剂和细菌以及转导噬菌体。
A method, which we calllocalized mutagenesis, is described for the isolation of temperature-sensitive and other types of mutations in any specific small region (about 1%) of the bacterial chromosome. The principle of this method is to mutate the transducing DNA rather than the bacterial DNA. One can select for the introduction of this mutated DNA into any particular region of the bacterial chromosome by transducing an auxotrophic marker in that region to prototrophy, thereby introducing new mutations in the neighborhood. We have used this method to isolate many different temperature-sensitive mutations in genes of unknown function in particular regions of the chromosome. Since the method is very simple, it can be used to saturate any region of the map with mutations in essential genes, or for various types of genetic manipulations. Although we have used hydroxylamine-mutagenized phage P22 andSalmonella typhimurium, the method should be applicable to other mutagens and bacteria and transducing phage.