Molecular cloning of growth hormone receptor (GHR) from common carp (Cyprinus carpio L.) and identification of its two forms of mRNA transcripts

The cDNA of growth hormone receptor (GHR) was cloned from the liver of 2-year common carp (Cyprinus carpio L.) by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE). Its open reading frame (ORF) of 1806 nucleotides is translated into a putative peptide of 602 amino acids, including an extracellular ligand-binding domain of 244 amino acids (aa), a single transmembrane domain of 24 aa and an intracellular signal-transduction domain of 334 aa. Sequence analysis indicated that common carp GHR is highly homologous to goldfish (Carassius auratus) GHR at both gene and protein levels. Using a pair of gene-specific primers, a GHR fragment was amplified from the cDNA of 2-year common carp, a 224 bp product was identified in liver and a 321 bp product in other tissues. The sequencing of the products and the partial genomic DNA indicated that the difference in product size was the result of a 97 bp intron that alternatively spliced. In addition, the 321 bp fragment could be amplified from all the tissues of 4-month common carp including liver, demonstrating the occurrence of the alternative splicing of this intron during the development of common carp. Moreover, a semi-quantitative RT-PCR was performed to analyze the expression level of GHR in tissues of 2-year common carp and 4-month common carp. The result revealed that in the tissues of gill, thymus and brain, the expression level of GHR in 2-year common carp was significantly tower than that of 4-month common carp.

Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in two freshwater shrimps, Palaemon modestus and Macrobrachium nipponensis, from a large shallow, eutrophic lake of the subtropical China

So far no information is available on microcystin (MC) contents in shrimps, prawns or crayfish from natural freshwaters. Tissue distributions and seasonal dynamics of the hepatotoxic MC-LR and -RR in two freshwater shrimps, Palaemon modestus and Macrobrachium nipponensis were studied monthly (during June-November, 2003) in a Chinese lake containing toxic cyanobacterial blooms. The shrimps P. modestus and M. nipponensis accumulated high MCs not only in the hepatopancreas (mean 4.29 and 0.53 mu g g(-1) DW, respectively) but also in the gonad (mean 1.17 and 0.48 mu g g-1 DW, respectively), and the crayfish Procambarus clarkii accumulated as much as 0.93 mu g g(-1) DW in the gonad. This indicates that gonads of these invertebrates are the second important target organ of MCs. P. modestus apparently accumulated more MCs in their organs than M. nipponensis, which might be a reflection of their difference in trophic niche. Eggs of the shrimps accumulated 8.4% (M. nipponensis, 0.27 mu g g(-1) DW) and 29.0% (P. modestus, 2.34 mu g g(-1) DW) of total toxin burden, indicating that MCs had been transferred into offspring from their adults. Among the shrimp muscle samples analyzed, 31% were above the provisional WHO TDI level, suggesting the risk of consuming shrimps in Lake Chaohu. (c) 2005 Elsevier Ltd. All rights reserved.

Characterization of two membrane-associated protease genes obtained from screening out-membrane protein genes of Flavobacterium columnare G(4)

In order to identify genes encoding the outer membrane proteins (OMPs) of the myxobacter Flavobacterium columnare G(4), the expression library of the bacterium was screened by using rabbit antisera developed against its OMPs. Positive colonies of Escherichia coli M15 containing fragments encoding the bacterial OMPs were selected for cloning the relevant genes by genomic walking methods. Two genes encoding a membrane-associated zinc metalloprotease and prolyl oligopeptidase are reported in this paper. The membrane-associated zinc metalloprotease gene (map) is 1800 bp in length, coding for 449 amino acids (aa). Despite the presence of a conserved motif HEXXH for all metalloproteases, the special HEXXH similar to 32 aa similar to E motif of the F. columnare G(4) Map and its low level of identity with other reported zinc-containing metalloproteases may imply that the membrane-associated zinc metalloprotease of F. columnare G(4) represents a new family of zincins. The gene encoding prolyl oligopeptidase (Pop), a serine proteinase, is 2352 bp in length, coding for 649 aa. Sequence homology analysis revealed that the Pop is also novel as it has <50% identity with other reported prolyl oligopeptidase family proteins. The present study represents the first to employ anti-fish bacterial OMP sera to screen genes of membrane-associated proteases of fish pathogenic bacteria, and to provide necessary information for the examination of the role of the two genes in the infection and pathogenesis of F. columnare.

Identification and expression analysis of two IFN-inducible genes in crucian carp (Carassius auratus L.)

Interferon (IFN) exerts its antiviral effects mainly through activation of a subset of IFN-stimulated genes (ISG), but relatively few of fish ISGs have been isolated and characterized so far. Here, we report two fish ISGs, termed CaIF158 and CaIF156, cloned from a subtractive cDNA library constructed with mRNAs obtained from crucian carp (Carassius auratus L.) blastulae embryonic (CAB) cells infected by UV-inactivated GCHV and mock-infected cells. Database search revealed that both ISGs had a high-level homology with all members of a well conserved gene family with multiple tetratricopeptide repeat (TPR) motifs, including human IF160, IF158, IF156, IFI54 and their homologues in some other mammalian species. The transcripts of CaIF158 and CaIF156 were undetectable in CAB cells but could be induced by active GCHV, UV-inactivated GCHV or CAB IFN. Analysis of expression difference between them and IFN signal factors, CaSTAT1 and CaIRF7, indicated that their transcriptions were mediated possibly through JAK-STAT signal pathway, which was further supported by the induction analysis in UV-inactivated GCHV infected, IFN-treated and untreated cells in the presence or absence of cycloheximide (CHX), a potent inhibitor of protein synthesis. In addition, a pufferfish (Fugu rubrides) DNA sequence representing putative FrIFI56 was also revealed when CalF158 and CalF156 were used to search the pufferfish genome database. Phylogenetic analysis showed that these fish ISGs form a unique clad independent of mammalian homologues, reflecting a distant evolutionary relationship from mammals. These studies identified the first teleost IFI56 and IFI58 orthologues. (C) 2003 Elsevier B.V All rights reserved.

Growth and feed utilization in two strains of gibel carp, Carassius auratus gibelio: paternal effects in a gynogenetic fish

Gibel carp (Carassius auratus gibelio Bloch) is a natural gynogenetic fish which requires sperm of the same or related species to activate egg development. The eggs of one gibel carp were divided into two batches. One batch was 'fertilized' with sperm from gibel carp (strain DD), and the other 'fertilized' with sperm from red common carp (Cyprinus carpio red variety) (strain DR). The juveniles were transferred to the laboratory 36 days post-hatch. Triplicate groups of each strain were fed a formulated diet at either 3% or satiation ration for 8 weeks. At both the restricted and satiation rations, specific growth rate was significantly higher in strain DR than in strain DD. At the 3% ration, there was no significant difference in feeding rate or feed conversion efficiency between the two strains. At the satiation ration, strain DR had a significantly lower feeding rate but higher feed conversion efficiency than strain DD. At the satiation ration, strain DR had a significantly lower intake protein, but higher recovered protein than strain DD. There was no significant difference in faecal protein loss between the two strains. At the 3% ration, strain had no significant effects on intake protein, faecal protein or recovered protein. Neither faecal energy loss nor recovered energy was affected by strain or ration. At both the 3% and satiation ration, final body contents of dry matter and lipid were significantly lower in strain DR than strain DD, while there was no significant difference in protein and energy content between the two strains at either ration level. The results suggested that gibel carp 'fertilized' with sperm of common carp grew faster than those 'fertilized' with sperm of gibel carp through increased feed conversion efficiency and protein retention.

RAPD and AFLP techniques for the analysis of genetic relationships in two genera of Decapoda

RAPD was used fur analysing three (sub-)species of mitten crabs (Eriocheir sinensis, E. japonicus, and E. japonicus hepuensis) and three populations of E. sinensis. The results show that their relationships on DNA level are similar to the classical taxonomic hypotheses (Dai, 1991). No diagnostic RAPD marker could be found, but there were statistically significant genetic differences among these taxa (P < 0.001) or populations (P < 0.001). That is, the intraspecific similarities were larger than the interspecific similarities; the intrasubspecific similarities were larger than the intraspecific similarities; and the intrapopulational similarities were larger than the interpopulational similarities. In AFLP analysis, no significant genetic difference has been found between E. sinensis and E. japonicus, but AFLP markers among four species of Macrobrachium (M. rosenbergii. M. nipponense, M. hainanense, and M. asperulum) were found. The DNA similarities among these four species of Macrobrachium are in accordance with morphological similarities.

Exact result for the Green's function of a two-dimensional interacting electron gas with spin-orbit interaction in a perpendicular magnetic field

An exact property is established for the Green's function of a uniform two-dimensional interacting electron gas in a perpendicular magnetic field with spin-orbit interaction. It is shown that the spin-diagonal Green's function is exactly diagonal in the Landau level index even in the presence of electron-electron interactions. For the Green's function with different spin indexes, only that with adjacent Landau level indexes is non-zero. This exact result should be helpful in calculating the Green's function approximately.

Investigation of filling factor in In0.53Ga0.47As/In0.52Al0.48As quantum wells with two occupied subbands

The magnetic field dependence of filling factors has been investigated on InP based In-0.53 Ga0.47As/In-0.52 Al-0.48 As quantum well samples with two occupied subbands by means of magnetotransport measurements at the temperature of 1.5 K in a magnetic field range of 0 to 13 T. Under the condiction that Laundau-level broadening is larger than the spin splitting of each subband, filling factors are even when the splitting energy of two subbands is an integer multiple of the cyclotron energy, i. e. Delta E-21 = khw(c). If the splitting energy of two subbands is half of an odd interger multiple of the cyclotron erergy, i. e. Delta E-21 = (2 k + 1) hw(c) /2, the filling factor is odd.

Quantum mechanical simulation of nanosized metal-oxide-semiconductor field-effect transistor using empirical pseudopotentials: A comparison for charge density occupation methods

The atomistic pseudopotential quantum mechanical calculations are used to study the transport in million atom nanosized metal-oxide-semiconductor field-effect transistors. In the charge self-consistent calculation, the quantum mechanical eigenstates of closed systems instead of scattering states of open systems are calculated. The question of how to use these eigenstates to simulate a nonequilibrium system, and how to calculate the electric currents, is addressed. Two methods to occupy the electron eigenstates to yield the charge density in a nonequilibrium condition are tested and compared. One is a partition method and another is a quasi-Fermi level method. Two methods are also used to evaluate the current: one uses the ballistic and tunneling current approximation, another uses the drift-diffusion method. (C) 2009 American Institute of Physics. [doi:10.1063/1.3248262]

Annealing ambient controlled deep defect formation in InP

Deep defects in annealed InP have been investigated by deep level transient capacitance spectroscopy (DLTS), photo induced current transient spectroscopy (PICTS) and thermally stimulated current spectroscopy (TSC). Both DLTS results of annealed semiconducting InP and PICTS and TSC results of annealed semi-insulating InP indicate that InP annealed in phosphorus ambient has five defects, while lid? annealed in iron phospbide ambient has two defects. Such a defect formation phenomenon is explained in terms of defect suppression by the iron atom diffusion process. The correlation of the defects and the nature of the defects in annealed InP are discussed based on the results.

Deep level defects in high temperature annealed InP

Deep level defects in high temperature annealed semi-conducting InP have been studied by deep level transient spectroscopy (DLTS). There is obvious difference in the deep defects between as-grown InP, InP annealed in phosphorus ambient and iron phosphide ambient, as far as their quantity and concentration are concerned. Only two defects at 0.24 and 0.64 eV can be detected in InP annealed in iron phosphide ambient, while defects at 0.24, 0.42, 0.54 and 0.64 eV have been detected in InP annealed in phosphorus ambient, in contrast to two defects at 0.49 and 0.64 eV or one defect at 0.13 eV in as-grown InP. A defect suppression phenomenon related to iron diffusion process has been observed. The formation mechanism and the nature of the defects have been discussed.

Generation of a supersinglet of three three-level atoms in cavity QED

We propose a scheme to generate a supersinglet of three three-level atoms in microwave cavity quantum electrodynamics based on the resonant atom-cavity interaction. In the scheme, three three-level atoms in suitable initial states are sequentially sent through three cavities originally prepared in their vacuum states. After an appropriate atom-cavity interaction process, in the subsequent measurement on the third cavity field the atoms are projected onto the desired supersinglet. The practical feasibility of this method is discussed.

Thermally induced Fe atom transition from substitutional to interstitial sites in InP and its influence on material property

As-grown Fe-doped semi-insulating InP single crystal has been converted into n-type low-resistance material after high temperature annealing. Defects in the InP materials have been studied by conventional Hall effect measurement, thermally stimulated current spectroscopy, deep level transient spectroscopy and X-ray diffraction respectively. The results indicate that Fe atoms in the InP material change from the substitutional to the interstitial sites under thermal activation. Consequently, the InP material loses its deep compensation centers which results in the change in types of conduction. The mechanism and cause of the phenomena have been analyzed through comparison of the sites of Fe atom occupation and activation in doping, diffusion and ion implantation processes of InP.

Optical properties of the E-0+Delta(0) energy level higher than the bandgap of GaAs studied by micro-photoluminescence technique

Using micro-photoluminescence technique, we observed a new photoluminescence peak about 0.348 eV above the bandgap of GaAs (E-0). By analyzing its optical characteristics, we assigned this peak to the nonequilibrium luminescence emission from the E-0 + Delta(0) bandgap in semi-insulated GaAs, which was further verified by Raman results. The observed polarization, excitation power dependence and temperature dependence of the photoluminescence spectra from the E-0 + Delta(0) energy level were very similar to those from the E-0 of GaAs. This mainly resulted from the common conduction band around Gamma(6) that was involved in the two optical transition processes, and indicated that the optical properties of bulk GaAs were mainly determined by the intrinsic properties of the conduction band. Our results demonstrated that the micro-photoluminescence technique is a powerful tool to investigate the high energy states above the fundamental bandgap in semiconductor materials.

Atomic configurations of dislocation core and twin boundaries in 3C-SiC studied by high-resolution electron microscopy

The defects in 3C-SiC film grown on (001) plane of Si substrate were studied using a 200 kV high-resolution electron microscope with point resolution of 0.2 nm. A posterior image processing technique, the image deconvolution, was utilized in combination with the image contrast analysis to distinguish atoms of Si from C distant from each other by 0.109 nm in the [110] projected image. The principle of the image processing technique utilized and the related image contrast theory is briefly presented. The procedures of transforming an experimental image that does not reflect the crystal structure intuitively into the structure map and of identifying Si and C atoms from the map are described. The atomic configurations for a 30 degrees partial dislocation and a microtwin have been derived at atomic level. It has been determined that the 30 degrees partial dislocation terminates in C atom and the segment of microtwin is sandwiched between two 180 degrees rotation twins. The corresponding stacking sequences are derived and atomic models are constructed according to the restored structure maps for both the 30 degrees partial dislocation and microtwin. Images were simulated based on the two models to affirm the above-mentioned results.