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2009年林道诺贝尔奖获得者会议:沃纳·阿尔伯,1978年诺贝尔生理学或医学奖获得者

The 2009 Lindau Nobel Laureate Meeting: Werner Arber, physiology or medicine 1978.

作者信息

Arber Werner

出版信息

J Vis Exp. 2010 Mar 10(37):1571. doi: 10.3791/1571.

Abstract

Swiss microbial geneticist, Werner Arber shared the 1978 Nobel Prize in Physiology or Medicine with Hamilton Smith and Daniel Nathans for their discovery of restriction endonucleases. Werner Arber was born in Granichen, Switzerland in 1929. Following a public school education, he entered the Swiss Polytechnical School in Zurich in 1949, working toward a diploma in natural sciences. There, his first research experience involved isolating and characterizing an isomer of chlorine. Following graduation in 1953, Arber joined a graduate program at the University of Geneva, taking on an assistanceship in electron microscopy (EM), in which he studied gene transfer in the bacterial virus (bacteriophage) lambda. Eventually encountering limitations with EM as a tool, he began using microbial genetics as a methodology for his studies. The study of microbial genetics had been possible for a relatively short time: DNA had been discovered to carry genetic information only a decade before he d entered the field. After earning his Ph.D. in 1958, Arber continued to develop skills in microbial genetics, working with colleagues in the United States for a short time before returning to Geneva at beginning of 1960. There, he continued working on lambda transduction in E. coli, but found that the virus would not efficiently propagate. Recalling research done seven years earlier by Joe Bertani and Jean Weigle on "host-controlled restriction-modification", he realized there must be a host-controlled modification of the invading DNA, and sought to identify the mechanism. Based on Grete Kallengerger s work that demonstrated degradation of both irradiated and non-irradiated phage lambda following injection in a host, Arber and his graduate student, Daisy Dussoix further investigated the fate of DNA, and found that restriction and modification (later determined to be postreplicative nuclotide methylation) directly affected DNA, but did not cause mutations. They also found that theses were properties of the bacterial strains, and that both viral and cellular DNA were degraded. Together, Arber and Dussoix reported their findings to scientific community in 1961 at the First International Biophysics Congress in Stockholm. Aber also presented the research to the Science Faculty of University of Geneva in 1962, earning the Plantamour-Prevost prize. Based on his work and the work of others, he hypothesized that an enzyme in the host bacterium cut DNA into smaller pieces at specific sites, and methylase modified the host DNA to protect it from the digestive enzyme. These theories were later confirmed by Urs Kuhnlein, who found that mutation of specific sites rendered the phage resistant to cleavage; Hamilton smith, who identified Type II endonuclease HindII; and Daniel Nathans, who used HindII to break the SV40 virus into 11 fragments, allowing him to determine its method of replication. Since the discovery of restriction endonucleases, researchers have used them as tools to study the functions of genes of all types of organisms. Restriction enzymes have also facilitated the study of gene functions and enabled production of substances of medical and nutritional importance. Arber feels that in the next few decades we will learn much from the study of epigentics --factors that can affect the phenotype of an organism without changing the genetic information--. He is proud that, in that studying restriction degradation and DNA methylation in the 1960s, he was among the first in studying epigenetic phenomenon.

摘要

瑞士微生物遗传学家沃纳·阿尔伯与汉密尔顿·史密斯和丹尼尔·内森斯共同获得了1978年诺贝尔生理学或医学奖,以表彰他们对限制性内切酶的发现。沃纳·阿尔伯于1929年出生在瑞士的格拉尼申。在接受完公立学校教育后,他于1949年进入苏黎世的瑞士联邦理工学院,攻读自然科学文凭。在那里,他的首次研究经历是分离和表征氯的一种异构体。1953年毕业后,阿尔伯加入了日内瓦大学的研究生项目,担任电子显微镜(EM)方面的助理,在其中他研究了细菌病毒(噬菌体)λ中的基因转移。最终,由于意识到电子显微镜作为一种工具存在局限性,他开始将微生物遗传学作为其研究方法。微生物遗传学的研究在当时开展的时间相对较短:就在他进入该领域的十年前,人们才发现DNA携带遗传信息。1958年获得博士学位后,阿尔伯继续提升微生物遗传学方面的技能,他在美国与同事合作了一段时间,然后于1960年初回到日内瓦。在那里,他继续研究大肠杆菌中的λ转导,但发现该病毒无法有效繁殖。回忆起七年前乔·贝尔塔尼和让·韦格勒关于“宿主控制的限制 - 修饰”的研究,他意识到入侵的DNA必定存在宿主控制的修饰,并试图确定其机制。基于格蕾特·卡伦格伯格的研究,该研究表明注射到宿主中的辐照和未辐照的噬菌体λ都会发生降解,阿尔伯和他的研究生黛西·迪苏瓦进一步研究了DNA的命运,发现限制和修饰(后来确定为复制后核苷酸甲基化)直接影响DNA,但不会导致突变。他们还发现这些是细菌菌株的特性,并且病毒和细胞DNA都会被降解。1961年,阿尔伯和迪苏瓦在斯德哥尔摩举行的第一届国际生物物理学大会上向科学界报告了他们的发现。阿尔伯还于1962年在日内瓦大学理学院展示了这项研究,获得了普兰塔穆尔 - 普雷沃斯特奖。基于他自己以及其他人的研究工作,他推测宿主细菌中的一种酶会在特定位点将DNA切割成更小的片段,而甲基化酶会修饰宿主DNA以保护其免受消化酶的作用。这些理论后来得到了乌尔丝·库恩莱因的证实,她发现特定位点的突变使噬菌体对切割产生抗性;汉密尔顿·史密斯鉴定出了II型内切酶HindII;丹尼尔·内森斯则使用HindII将SV40病毒切割成11个片段,从而能够确定其复制方式。自从发现限制性内切酶以来,研究人员一直将它们用作研究各类生物体基因功能的工具。限制性酶还推动了基因功能的研究,并使具有医学和营养重要性的物质的生产成为可能。阿尔伯认为,在未来几十年里,我们将从表观遗传学的研究中学到很多东西——表观遗传学是指那些能够在不改变遗传信息的情况下影响生物体表型的因素。他感到自豪的是,在20世纪60年代研究限制降解和DNA甲基化时,他是最早研究表观遗传现象的人之一。

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