My Life with Bacteriophage φ29

Abstract

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 29 for 45 years has been very rewarding. I can say that I was lucky because I did not expect that 29 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 29 DNA transcription and, in particular, the finding for the first time of a protein covalently linked to the 5-ends of 29 DNA that we later showed to be the primer for the initiation of phage DNA replication. Very relevant was the discovery of the 29 DNA polymerase, with its properties of extremely high processivity and strand displacement capacity, together with its high fidelity. The 29 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. Early Years

Target-Induced Nano-Enzyme Reactor Mediated Hole-Trapping for High-Throughput Immunoassay Based on a Split-Type Photoelectrochemical Detection Strategy

“…17 Herein we present a novel split-type photoelectrochemical immunoassay (STPIA) method for ultrasensitive and high-throughput detection of prostate-specific antigen (PSA, as a model analyte). To achieve this design, RCA-triggered formation of enzymatic concatamers on the nanogold particle was coupled with enzymatic hydrolysate-enhanced photocurrent response of CdS quantum dot (QD)-modified TiO 2 nanotube array (Scheme 1).…”

Taxonomy of prokaryotic viruses: update from the ICTV bacterial and archaeal viruses subcommittee

“…Consequently, the Subcommittee decided that, unless there was sufficient historical precedent (e.g., U29 or UX174), Phi would no longer be added to genus names. In addition, Greek letters can create problems in electronic databases, as exemplified by a PubMed search for references on Bacillus phage U29 [1], which retrieved articles on phi 29, phi29, Phi 29, Phi29, 29 phi, {phi}29, u29, and u29 phages. Therefore, the Subcommittee strongly discourages phage scientists from using Phi or any other Greek letter in virus and virus taxon names in the future; 4. elimination of hyphens from taxon names.…”

What can we learn from the construction of in vitro replication systems?

“…Most of the aforementioned replication systems are artificially designed; no equivalent system exists in nature, except for the RNA replication system described above. Attempts to reproduce natural viral replication systems have also been reported, such as the genome replication systems of bacteriophages phi29 and T4, adeno‐associated virus, norovirus, SV40, and influenza virus . In 1984, the more complex replication of the Escherichia coli genome was reconstituted in vitro from nine components by Kornberg's group .…”