Methylation and gene mutation in eukaryotic DNA

5-methylcytosine (m(5)C) as a rare base exists in eukaryotic genomes, which is a normal constitution in many eukaryotic DNA and the existence of m(5)C is a feature of eukaryotic DNA. Under regular physiological conditions, cytosine of eukaryotic DNA is usually methylated. Up to the present, many people consider that the m(5)C may be mutation hotspots by the deamination leading to gene mutation. Our study indicated that the spontaneous mutation caused by the transition of G.C --> A.T, in eukaryotic DNA, may result from the tautomer changing of base pairs and may also be cause by other factor actions, however it could not be caused by the deamination of m(5)C.

mtDNA haplogroup distribution in Chinese patients with Leber's hereditary optic neuropathy and G11778A mutation

Mitochondrial DNA background has been shown to be involved in the penetrance of Leber's hereditary optic neuropathy (LHON) in western Eurasian populations. To analyze mtDNA haplogroup distribution pattern in Han Chinese patients with LHON and G11778A muta

Mitochondrial DNA mutation m.3635G > A may be associated with Leber hereditary optic neuropathy in Chinese

Leber hereditary optic neuropathy (LHON) was the first disease to be linked to the presence of a mitochondrial DNA (mtDNA) mutation. Nowadays over 95% of LHON cases are known to be caused by one of three primary mutations (m.11778G>A, m.14484T>C, and m.34

Gene mapping of a new coat mutation in mouse

Gene mapping of a mouse coat mutation has been investigated. First, 100 10-bp random primers were used to amplify DNA, but the mutation could not be located by this method because there were no correlation between the amplified products and coat phenotypes. Second, by using Idh1, Car2, Mup1, Pgb1, Hbb, Es10, Es1, Mod1, Gdc1, Ce2, Es3 as genetic markers, linkage test crosses (two-point test) consisting of intercrossing uncovered BALB/c mice (homozygotes) to CBA/N and C57BL/6 mice with normal hair and backcrossing the heterozygotes of the F1 to the uncovered BALB/c mice were made. It was soon evident that the mutation was linked to Es3 on chromosome 11. Furthermore, three-point test was made by using Es3 and D11Mit8 (a microsatellite DNA) as genetic markers. The result showed that the mutation was linked to Es3 with the percentage recombination of (7.89 +/- 2.19)%, and linked to D11Mit8 with the percentage recombination of (26.38 +/- 3.57)%. The percentage recombination between Es3 and D11Mit8 was (32.90 +/- 3.81)%. The mutation was named Uncovered, with the symbol Uncv. According to the recombinations, the loci order was D11Mit8-26.30 +/- 3.57- Uncv-7.89 +/- 2.19-Es3. From the location on the chromosome, it was concluded that the mutation was a new mutation which affected the skin and hair structure of mouse. The Uncv has entered MGD (Mouse Genome Database).

Designing output-feedback predictive controllers by reverse-engineering existing LTI controllers

An approach to designing a constrained output-feedback predictive controller that has the same small-signal properties as a pre-existing output-feedback linear time invariant controller is proposed. Systematic guidelines are proposed to select an appropriate (non-unique) realization of the resulting state observer. A method is proposed to transform a class of offset-free reference tracking controllers into the combination of an observer, steady-state target calculator and predictive controller. The procedure is demonstrated with a numerical example. © 2013 IEEE.

Pressure-assisted field-amplified sample injection with reverse migrating micelles for analyzing trace steroids in MEKC

This paper describes a novel method that applies pressure-assisted field-amplified sample injection with reverse migrating micelles (PA-FASI-RMM) for the online concentration of neutral analytes in MEKC with a low-pH BGE. After injection of a plug of water into the separation capillary, negative voltage and positive pressure were simultaneously applied to initialize PA-FASI-RMM injection. The hydrodynamic flow generated by the positive pressure compensated the reverse EOF in the water plug and allowed the water plug to remain in the capillary during FASI with reverse migrating micelles (FASI-RMM) to obtain a much longer injection time than usual, which improved stacking efficiency greatly. Equations describing this injection mode were introduced and were supported by experimental results. For a 450-s online PA-FASI-RMM injection, three orders of magnitude sample enhancement in terms of peak area could be observed for the steroids and an achievement of detection limits was between 1 and 10 ng/mL.

Vertical variation in dissolved alkaline phosphatase activity in a shallow eutrophic lake determined in reverse micelles

We evaluated the feasibility of microencapsulating dissolved alkaline phosphatase of a water body into reverse micelle systems prepared by hexadecyltrimethylammonium bromide as a surfactant in cyclohexane and 1-butanol as co-surfactant. The dissolved alkaline phosphatase activity within the micelle was described, including its kinetic parameters and the effects of pH and temperature on catalytic activity in surface, overlying and interstitial water of Lake Donghu. We found the similarities on the behavior of dissolved alkaline phosphatase of surface and interstitial water in reverse micelles, which was distinctly different from its behavior in the overlying water. This difference likely reflected the different origins of the dissolved alkaline phosphatase in the vertical profile of the lake. This system provides a novel tool with which to study the diversity and ecological significance of extracellular enzymes in aquatic environments.

Complex mutation at a microsatellite locus in sturgeons: Acipenser sinensis, A-schrenckii, A-gueldenstaedtii and A-baerii

Cross-species amplifications of microsatellite locus Spl-106, which was originally screened from the genome of shovelnose sturgeon (Scaphirhynchus platorynchus) with a perfect TAGA repeat motif, were carried out in four other species of the genera Acipenser. A total of 34 polymerase chain reaction (PCR) products representing 16 different alleles of this locus was sequenced. Sequence analysis results showed that besides the number changes of repeat units, many mutational events, such as single-base substitutions and various insertion/deletion (indels) occurred not only at species level but also at individual level, even among the different alleles within the same individual. The repeat motifs varied from perfect (TAGA)n array to perfect compound (TAAA)m (GAAA)n and perfect or imperfect compound (TAAA)m (TAGA)n (TAAA)x arrays in different species and different individuals. The evolution dynamics of this locus in sturgeons was inferred in that it may evolve from a single perfect to different perfect or imperfect compounds.

A detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription polymerase chain reaction

A rapid, sensitive and highly specific detection method for grass carp hemorrhagic virus (GCHV) based on a reverse transcription-polymerase chain reaction (RT-PCR) has been developed. Two pairs of PCR primers were synthesized according to the cloned cDNA sequences of the GCHV-861 strain. For each primer combination, only one specific major product was obtained when amplification was performed by using the genomic dsRNA of GCHV-861 strain. The lengths of their expected products were 320 and 223 bp, respectively. No products were obtained when nucleic acids other than GCHV-861 genomic RNA were used as RT-PCR templates. To assess the sensitivity of the method, dilutions of purified GCHV-861 dsRNA total genome (0.01 pg up to 1000 pg) were amplified and quantities of as little as 0.1 pg of purified dsRNA were detectable when the amplification product was analyzed by 1.5% agarose gel electrophoresis. This technique could detect GCHV-861 not only in infected cell culture fluids, but also in infected grass carp Ctenopharyngodon idellus and rare minnow Gobiocypris rarus with or without hemorrhagic symptoms. The results show that the RT-PCR amplification method is useful for the direct detection of GCHV.

Influence of indium-tin-oxide thin-film quality on reverse leakage current of indium-tin-oxide/n-GaN Schottky contacts

Indium-tin-oxide (ITO)/n-GaN Schottky contacts were prepared by e-beam evaporation at 200 degrees C under various partial pressures of oxygen. X-ray photoemission spectroscopy and positron beam measurements were employed to obtain chemical and structural information of the deposited ITO films. The results indicated that the observed variation in the reverse leakage current of the Schottky contact and the optical transmittance of the ITO films were strongly dependent on the quality of the ITO film. The high concentration of point defects at the ITO-GaN interface is suggested to be responsible for the large observed leakage current of the ITO/n-GaN Schottky contacts. (c) 2006 American Institute of Physics.

Study on the reverse characteristics of Ti/6H-SiC Schottky contacts

The reverse I(V) measurement and analytic calculation of the electron transport across a Ti/6H-SiC Schottky barrier are presented. Based on the consideration of the barrier fluctuations and the barrier height shift caused by image charge and the applied voltage drop across Ti/SiC interfical layer, a comprehensive analytical model for the reverse tunneling current is developed using a WKB calculation of the tunneling probability through a reverse biased Schottky barrier. This model takes into account the main reverse conduction mechanism, such as field emission, thermionic field emission and thermionic emission. The fact that the simulated results are in good agreement with the experimental data indicates that the barrier height shift and barrier fluctuation can lead to reverse current densities orders of magnitude higher than that obtained from a simple theory. It is shown that the field and thermionic field emission processes, in which carries can tunnel through the barrier but cannot surmount it with insufficient thermal energy, dominate the reverse characteristics of a SiC Schottky contacts in a normal working condition.

Influence of crystal perfection on the reverse leakage current of the SiGe Si p-n heterojunction diodes

Double-crystal X-ray diffraction and I-V characterization have been carried out on the GSMBE grown SiGe/Si p-n heterojunction materials. Results show that the SiGe alloys crystalline quality and the misfit dislocations are critical influences on the reverse leakage current. The crystal perfection and/or the degree of metastability of the Sice alloys have been estimated in terms of the model proposed by Tsao with the experimental results. High-quality p-n heterojunction diodes can be obtained by optimizing the SiGe alloy structures, which limit the alloys in the metastable states. (C) 1999 Elsevier Science B.V. All rights reserved.