Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid), Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain.
J Biol Chem. 2012 Dec 28;287(53):44568-79. doi: 10.1074/jbc.X112.433458. Epub 2012 Nov 2.
This article is a survey of my scientific work over 52 years. During my postdoctoral stay in Severo Ochoa's laboratory, I determined the direction of reading of the genetic message, and I discovered two proteins that I showed to be involved in the initiation of protein synthesis. The work I have done in Spain with bacteriophage ϕ29 for 45 years has been very rewarding. I can say that I was lucky because I did not expect that ϕ29 would give so many interesting results, but I worked hard, with a lot of dedication and enthusiasm, and I was there when the luck arrived. I would like to emphasize our work on the control of ϕ29 DNA transcription and, in particular, the finding for the first time of a protein covalently linked to the 5'-ends of ϕ29 DNA that we later showed to be the primer for the initiation of phage DNA replication. Very relevant was the discovery of the ϕ29 DNA polymerase, with its properties of extremely high processivity and strand displacement capacity, together with its high fidelity. The ϕ29 DNA polymerase has become an ideal enzyme for DNA amplification, both rolling-circle and whole-genome linear amplification. I am also very proud of the many brilliant students and collaborators with whom I have worked over the years and who have become excellent scientists. This Reflections article is not intended to be the end of my scientific career. I expect to work for many years to come.
这是一篇综述性文章,总结了我 52 年来的科研工作。在Severo Ochoa 的实验室做博士后期间,我确定了遗传信息的读取方向,并发现了两种参与蛋白质合成起始的蛋白质。我在西班牙用噬菌体 ϕ29 进行了 45 年的研究,收获颇丰。可以说我很幸运,因为我没想到 ϕ29 会带来如此多有趣的结果,但我努力工作,充满热情和奉献精神,幸运来临时我就在那里。我想强调一下我们在控制 ϕ29 DNA 转录方面的工作,特别是首次发现一种与 ϕ29 DNA 5'-末端共价连接的蛋白质,我们后来发现它是噬菌体 DNA 复制起始的引物。发现 ϕ29 DNA 聚合酶也非常重要,它具有极高的持续性和链置换能力,以及高度的保真度。ϕ29 DNA 聚合酶已成为滚环和全基因组线性扩增的理想酶。多年来,我与许多杰出的学生和合作者一起工作,他们已经成为优秀的科学家,我也为此感到非常自豪。这篇反思文章并不是我科学生涯的终点,我期望在未来的许多年里继续工作。