RESUMEN
Here, we present pRH030, a new CRISPR-Cas9 tool for the genetic engineering of Bacillus phages and beyond. It is based on the Streptococcus pyogenes cas9 with its native constitutive promoter, tracrRNA, and a gRNA precursor. The constitutive expression of Cas9 was conducive to the inactivation of viral attackers and enhanced phage mutagenesis efficiency up to 100%. The gRNA precursor can be built up to an artificial CRISPR array with up to 5 spacers (target sequences) assembled from ordinary oligonucleotides and directly cloned into pRH030. Required time and resources remain comparable to a single gRNA cloning. These properties make pRH030 an attractive new system for the modification of Bacillus phages and qualify it for research beyond genetic construction.
Asunto(s)
Fagos de Bacillus/genética , Bacillus subtilis/virología , Sistemas CRISPR-Cas , Fagos de Bacillus/fisiología , Ingeniería Genética , Mutagénesis , ARN Guía de Kinetoplastida/genética , ARN Guía de Kinetoplastida/metabolismoRESUMEN
We introduce a novel method for local structure determination with a spatial resolution of the order of 0.01 Å. It can be applied to materials containing clusters of exchange-coupled magnetic atoms. We use neutron spectroscopy to probe the energies of the cluster excitations which are determined by the interatomic coupling strength J. Since for most materials J is related to the interatomic distance R through a linear relation dJ/dR=α (for dR/Râª1), we can directly derive the local distance R from the observed excitation energies. This is exemplified for the mixed one-dimensional paramagnetic compound CsMn(x)Mg(1-x)Br3 (x=0.05,0.10) containing manganese dimers oriented along the hexagonal c axis. Surprisingly, the resulting Mn-Mn distances R do not vary continuously with increasing internal pressure but lock in at some discrete values.
RESUMEN
α-MnMoO(4) is a tetrameric magnetic cluster system which undergoes a transition to three-dimensional antiferromagnetic order at T(N) = 10.7 K. In the ordered state the Mn(2+) spins (s(ν) = 5/2) are ferromagnetically aligned within the tetramer, resulting in a total cluster spin S = 10. The magnetic excitation spectrum consists of eight excitation modes of tetramer origin propagating in the whole reciprocal space. From single-crystal inelastic neutron scattering investigations the three-dimensional coupling scheme is rationalized in the framework of a tetramer-based dispersion model. The tetramer states and the energy dispersion of all the magnetic excitations are described by Heisenberg-like intra- and intercluster exchange interactions, respectively, thus α-MnMoO(4) is a suitable tetrameric model system to study the interplay of these interactions.
RESUMEN
The oxygen isotope effect on the relaxation rate of crystal-field excitations in the slightly underdoped high-temperature superconductor HoBa2Cu4O8 has been investigated by means of inelastic neutron scattering. For the 16O compound there is clear evidence for the opening of an electronic gap in the normal state at T(*) approximately 170 K far above T(c) = 79 K. Upon oxygen isotope substitution ( 16O vs 18O) T(c) decreases marginally to 78.5 K, whereas T(*) is shifted to about 220 K. This huge isotope shift observed for T(*) which is absent in NMR and NQR experiments suggests that the mechanism leading to an isotope effect on the pseudogap has to involve a time scale in the range 10(-8)>>tau>10(-13) s.
RESUMEN
Inelastic neutron scattering was employed to study photoeffects on the molecular dynamics of membranes of the photosynthetic bacterium Rhodopseudomonas viridis. The main photoactive parts of this biomolecular system are the chlorophyll molecules whose dynamics were found to be affected under illumination by visible light in a twofold manner. First, vibrational modes are excited at energies of 12(2) and 88(21) cm(-1). Second, a partial "freezing" of rotational modes is observed at energies of 1.2(3) and 2.9(5) cm(-1). These results are attributed to a possible coupling between molecular motions and particular mechanisms in the photosynthetic process.