资源环境科技发展态势分析平台

Reverse genetics has been an indispensable tool to gain insights into viral pathogenesis and vaccine development. The genomes of large RNA viruses, such as those from coronaviruses, are cumbersome to clone and manipulate inEscherichia coliowing to the size and occasional instability of the genome(1-3). Therefore, an alternative rapid and robust reverse-genetics platform for RNA viruses would benefit the research community. Here we show the full functionality of a yeast-based synthetic genomics platform to genetically reconstruct diverse RNA viruses, including members of theCoronaviridae,FlaviviridaeandPneumoviridaefamilies. Viral subgenomic fragments were generated using viral isolates, cloned viral DNA, clinical samples or synthetic DNA, and these fragments were then reassembled in one step inSaccharomyces cerevisiaeusing transformation-associated recombination cloning to maintain the genome as a yeast artificial chromosome. T7 RNA polymerase was then used to generate infectious RNA to rescue viable virus. Using this platform, we were able to engineer and generate chemically synthesized clones of the virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)(4), which has caused the recent pandemic of coronavirus disease (COVID-19), in only a week after receipt of the synthetic DNA fragments. The technical advance that we describe here facilitates rapid responses to emerging viruses as it enables the real-time generation and functional characterization of evolving RNA virus variants during an outbreak.

Charged pions(1) are the lightest and longest-lived mesons. Mesonic atoms are formed when an orbital electron in an atom is replaced by a negatively charged meson. Laser spectroscopy of these atoms should permit the mass and other properties of the meson to be determined with high precision and could place upper limits on exotic forces involving mesons (as has been done in other experiments on antiprotons(2-9)). Determining the mass of the pi(-) meson in particular could help to place direct experimental constraints on the mass of the muon antineutrino(10-13). However, laser excitations of mesonic atoms have not been previously achieved because of the small number of atoms that can be synthesized and their typically short (less than one picosecond) lifetimes against absorption of the mesons into the nuclei(1). Metastable pionic helium (pi He-4(+)) is a hypothetical(14-16) three-body atom composed of a helium-4 nucleus, an electron and a pi(-) occupying a Rydberg state of large principal (n approximate to 16) and orbital angular momentum (l approximate to n - 1) quantum numbers. The pi He-4(+) atom is predicted to have an anomalously long nanosecond-scale lifetime, which could allow laser spectroscopy to be carried out(17). Its atomic structure is unique owing to the absence of hyperfine interactions(18,19) between the spin-0 pi(-) and the He-4 nucleus. Here we synthesize pi He-4(+) in a superfluid-helium target and excite the transition (n, l) = (17, 16) -> (17, 15) of the pi(-)-occupied pi He-4(+) orbital at a near-infrared resonance frequency of 183,760 gigahertz. The laser initiates electromagnetic cascade processes that end with the nucleus absorbing the pi(-) and undergoing fission(20,21). The detection of emerging neutron, proton and deuteron fragments signals the laser-induced resonance in the atom, thereby confirming the presence of pi He-4(+). This work enables the use of the experimental techniques of quantum optics to study a meson.

Long-lived pionic helium atoms (composed of a helium-4 nucleus, an electron and a negatively charged pion) are synthesized in a superfluid-helium target, as confirmed by laser spectroscopy involving the pion-occupied orbitals.