Chaos and order in spontaneous mutation.
Chaos and order in spontaneous mutation.
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自发突变中的混沌与秩序。
DOI：
10.1093/genetics/173.1.1
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发表时间：
2006
期刊：
影响因子：
3.3
通讯作者：
Drake,JohnW
中科院分区：
文献类型：
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作者：
Drake,JohnW
大约在1960年，我在伊利诺伊大学厄巴纳分校工作，研究纽卡斯尔病病毒（NDV），这是一种易于控制的动物病毒，感染的是鸡而不是人。（我以前研究过脊髓灰质炎病毒，对这三种病毒都获得了极高的血清滴度。）NDV空斑试验要求收获正确年龄的鸡胚并解离成单细胞，然后将其用于产生在几天内融合的单层。病毒样品被吸附到单层上，然后再孵育几天，同时形成噬斑。这个方案很繁琐，而且有一天的时间，结果是，这些时间并没有完全被实验的计划和分析、演讲的准备和发表以及一个底层学者的各种职责所填满，作为一名研究生，我一直关注着加州理工学院噬菌体小组的滑稽动作，也开始意识到诱变碱基类似物5-溴尿嘧啶。在接下来的几年里，恩斯特·弗里斯（Ernst Freese）和西摩·本泽（Seymour Benzer）利用噬菌体T4进行的早期突变研究给我留下了越来越深刻的印象。恩斯特特别研究了具有高度特异性作用的诱变剂的突变途径，例如，碱基类似物5-溴尿嘧啶和2-氨基嘌呤可诱导两个方向的转换，而羟胺仅诱导GÁC/AÁT转换。我经常有机会用紫外线照射NDV，我想知道紫外线照射的诱变特异性是否可以通过用碱基类似物、羟胺和原黄素对紫外线诱导的突变进行回复突变试验来确定，最后一种的特异性已经由Benzer和Sydney Brenner证明。从Raymond Latarjet为噬菌体T4开发的方案开始，我开始将噬菌体实验插入到自由日中，
Around 1960 I was working at the University of Illinois at Urbana using Newcastle disease virus (NDV), a tractable animal virus that infects chickens rather than humans.(Working previously with polioviruses, I had acquired extraordinarily high serum titers against all three strains.) The NDV plaque assay required that chicken embryos of the correct age be harvested and dissociated into single cells, which were then used to produce monolayers that became confluent in a few days. Virus samples were adsorbed to the monolayers, which were then incubated for a few more days while plaques formed. The regimen was cumbersome and contained day-long gaps that, as it turned out, were incompletely filled with the planning and analysis of experiments, the preparation and delivery of lectures, and the miscellaneous duties of a bottom-rung academic.As a graduate student I had followed the antics of the Caltech phage group and had also become aware of the mutagenic base analog 5-bromouracil. Over the next few years I was increasingly impressed by the early work on mutation by Ernst Freese and Seymour Benzer using phage T4. Ernst in particular was dissecting out mutational pathways with mutagens that seemed to be highly specific in their actions, for instance, the base analogs 5-bromouracil and 2-aminopurine that induced transitions in both directions and hydroxylamine that induced only GÁC/AÁT transitions. I often had occasion to UV irradiate NDV, and I wondered if the mutagenic specificity of UV irradiation could be ascertained by performing reversion tests on UV-induced mutations with base analogs, hydroxylamine, and proflavin, the specificity of the last having been demonstrated by Benzer and Sydney Brenner. Starting with a protocol developed by Raymond Latarjet for phage T4, I began to intercalate phage experiments into the free days pro-