Intracellular localization of PNA in human cells upon its introduction by electroporation

Peptide nucleic acid (PNA) is one of the most useful DNA analogs in a wide variety of gene analysis in human cells. In order to exhibit its maximal functions, PNA must be localized to a desired place (e.g., nucleus, cytoplasm and other organelles). Here, we introduced PNAs into HeLa cells by electroporation and examined their localization at various time points. The PNA which binds to the mitochondrial COII gene was initially accumulated in the nucleus, and thereafter mostly transferred to cytoplasm. This time-dependent intracellular localization of PNA is ascribed to the breakdown of the nuclear envelope in the cell division. On the other hand, another PNA that binds to telomere repeat sequence mostly remained in the nucleus, even after the cell division occurred. The retention of this PNA in the nucleus was further enhanced when it was conjugated with Cy3.     

Enzyme treatment-free and ligation-independent cloning using caged primers in polymerase chain reactions

A new simple scheme for constructing recombinant vectors that does not require any restriction enzyme, ligase, or any other special enzyme treatment has been developed. By using caged primers in PCR, unnatural sticky-ends of any sequence, which are sufficiently long for ligation-independent cloning (LIC), are directly prepared on the product after a brief UVA irradiation. Target genes and vectors amplified by this light-assisted cohesive-ending (LACE) PCR join together in the desired arrangement in a simple mixture of them, tightly enough to be repaired and ligated in competent cells.     

Polymerization of coordinated monomers. XII. Alternating copolymerization of γ-crotonolactone with styrene with the use of stannic chloride

γ-Crotonolactone and styrene copolymerize alternately in the presence of stannic chloride at -10°C under photoirradiation. The intrinsic viscosity of the resulting copolymer is in the range of 0.6–0.8 dl/g at 30°C in chloroform. The equilibrium constants for the complex formation between stannic chloride and γ-crotonolactone were determined in 1,2-dichloroethane-toluene solution at 0 and ?20°C by use of absorption band at 350 nm. Continuous variation plots based on the ¹H-chemical shift show a 1:1 interaction between styrene and the γ-crotonolactone coordinated to stannic chloride. The equilibrium constants for the ternary molecular complex formation between the coordinated γ-crotonolactone and styrene were determined in 1,2-dichloroethane in the temperature range from ?20 to 0°C. The equilibrium constants, derived independently from the measurements of the nonequivalent protons in γ-crotonolactone, are equal to each other within the experimental error. The mechanism of the alternating copolymerization of γ-crotonolactone and styrene in the presence of stannic chloride is discussed in terms of the homopolymerization of the ternary molecular complex.     

The interaction of dansyl amino acids with bovine serum albumin

The interaction of 5-dimethylaminonaphthalene-1-sulfonyl (DNS or dansyl) amino acids with bovine serum albumin (BSA) was investigated by means of fluorescence measurements. Fluorometric titrations revealed that BSA has one high affinity site (binding constant,K _a=10⁵10⁶ M^–1), and other sites of lower affinity (K _a=10³10⁴ M^–1) for the probes. Static excitation and emission spectra, lifetimes, time resolved emission spectra, and anisotropy data indicated that the binding is stabilized mainly through fixation by the high affinity binding site. The binding constant significantly decreased with the increase of the spacer distance between the dansyl and anionic groups of the probe molecule. This observation was explained by considering the change of the electrostatic interaction between the anionic group of the probe and a cationic residue in the vicinity of the site.     

Selective transport of D,L‐tryptophan through poly(L‐glutamic acid) membranes

Permeability coefficients of D ‐ and L ‐tryptophan (D ‐, L ‐Trp) were estimated for poly(L ‐glutamic acid) (PLG) membranes immersed in aqueous ethanol. D ‐tryptophan was selectively transported (the maximum permeability ratio was 2.6) depending on the amount and the species of crosslinking agent, and on the composition of immersing solvent. It is suggested that hydrogen bonding between uncharged permeates and carboxyl and/or amide groups of PLG is an essential factor for the selective transport. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 1035–1041, 1999     

THz imaging of cyclotron emission in quantum Hall conductors

Recent studies of cyclotron emission microscopy on quantum Hall related states are reported. The topics include non-equilibrium between edge and bulk states, current-induced breakdown of the quantum Hall effect, and the emission threshold at hot spots. Experimental method of scanning-type terahertz microscopes developed towards photon-counting level sensitivity is also described.     

Dynamic behavior of diffusion flame interacting with a large-scale vortex by laser imaging techniques

Instantaneous and simultaneous measurements of two-dimensional temperature and OH-LIF profiles by combining Rayleigh scattering with laser induced fluorescence (LIF) were demonstrated in a nitrogen-diluted hydrogen (H₂ 30% + N₂ 70%) laminar normal diffusion flame interacting with a large-scale vortex by oscillating central fuel flow or in an inverse diffusion flame by oscillating central airflow. The dynamic behavior of the diffusion flame extinction and reignition during the flame–vortex interaction processes was investigated. The results obtained are described as follows. (1) The width of the reaction zone decreases remarkably, and a decrease in flame temperature and OH-LIF is seen with increasing central airflow in an inverse diffusion flame. OH-LIF increases, and temperature does not change with increasing central fuel flow in a normal diffusion flame. The computations predict the experimental results well, and it is revealed that flame temperature characteristics result from the preferential diffusion of heat and species, which induces excess enthalpy or on enthalpy deficit, and an increase or decrease in H₂ mole fraction in the flame. (2) When a large velocity fluctuation is given to the central flow, the temperature and the OH-LIF at the reaction zone become thin at the convex and circumferential part of the vortex where a high temperature layer exists, and the temperature at the reaction zone is lowered in the inverse flame and the normal flame. (3) The width and temperature of the reaction zone interacting with the vortex recover quickly to that of the laminar steady flame after the vortex passing in the normal flame, but the recovery to that of the steady flame after the vortex passing is delayed in the inverse flame. (4) When a remarkably large velocity fluctuation is given to the central airflow in the inverse flame, thinning of temperature and reaction zone starts at the convex and circumferential part of the vortex, resulting in a and flame extinction completely occurs at the tail part of the vortex and makes the pair of edge flames. The outside edge flame reignites and connects with the upstream reaction zone. The inside edge flame finally extinguishes as the supply of fuel is interrupted by the outside edge flame.     

One‐Pot Substitution‐Rearrangement Reaction of 2‐Chloroglycidic Ester: Effective Protocol for the Preparation of 3‐Arylsulfenyl‐2‐keto Ester

The one‐pot substitution‐rearrangement reaction of 2‐chloroglycidic ester is reported. Treatment of 2‐chloroglycidic ester with sodium arylthiolate at room temperature results in the formation of 3‐arylsulfenyl‐2‐keto ester in excellent to good yields.     

A capacitive method for hall effect measurements

A new, simple and reliable method for Hall effect measurements is introduced. The method employs a capacitive probe technique and requires neither special shaping of samples nor probing Hall contacts. With this method, Hall effect measurements onp-Ge have been first extended to high electric fields up to 3kV/cm at 4.2 K. The wide applicability of this method is discussed.     

Parallel fabrication of artificially designed superlattices by combinatorial laser MBE

A system for parallel synthesis of atomically regulated lattices and nano-structured heterojunctions has been developed. This combinatorial laser MBE system is composed of a UHV-PLD chamber, scanning RHEED for in situ diagnostics of reactions at various areas of substrate surface, a pair of fixed and moving masks to define the film deposition area, and a computer for controlling the operation conditions as functions of deposition area and time. In this system, oxide layered lattice strips were deposited in parallel on a SrTiO₃ substrate under RHEED observation of individual strips with different compositions and/or layer thickness.