Detection of potential positive regulatory motifs of transcription in yeast introns by comparative analysis of oligonucleotide frequencies

We conducted a comparative statistical analysis of tetra- through hexanucleotide frequencies in two sets of introns of yeast genes. The first set consisted of introns of genes that have transcription rates higher than 30 mRNAs/h while the second set contained introns of genes whose transcription rates were lower than or equal to 10 mRNAs/h. Some oligonucleotides whose occurrence frequencies in the first set of introns are significantly higher than those in the second set of introns were detected. The frequencies of occurrence of most of these detected oligonucleotides are also significantly higher than those in the exons flanking the introns of the first set. Interestingly some of these detected oligonucleotides are the same as well known "signature" sequences of transcriptional regulatory elements. This could imply the existence of potential positive regulatory motifs of transcription in yeast introns. (C) 2003 Elsevier Ltd. All rights reserved.

Molecular systematics of medusae in the genus Craspedacusta (Cnidaria: Hydrozoa: Limnomedusae) in China with the reference to the identity of species

The objective of this study was to illustrate the phylogenetic relationship of the species in the genus Craspedacusta in China. The medusae samples were collected at 28 localities in China representing seven described species with their entire ITS region (the contiguous sequences of ITS-1, 5.8S and ITS-2 rDNA) rDNA sequences cloned. Among the 28 samples, the range of sequence variation in the complete ITS and 5.8S region was between 0 and 36.2%. Three main clades were revealed by both maximum likelihood and neighbour-joining trees, with sequence difference of 0-0.9, 0-3.7 and 0.1-1.5% in the three clades. The nesting of C. xinyangensis representatives within C. sowerbii, C. brevinema within C. sinensis and C. sichuanensis within C. kiatingi is strongly supported, with interspecific sequence divergence of 0-0.9, 0.1-1.4 and 0.0-0.4%, respectively. Thus, it is suggested that C. xinyangensis should be the synonym of C. sowerbii, C. sichuanensis the synonym of C. kiatingi and C. brevinema the synonym of C. sinensis. However, the taxonomic status of C. ziguiensis is still uncertain. According to the tree topology, C. kiatingi was closer to C. sowerbii than to C. sinensis. Craspedacusta sinensis was the most genetically distinct from distance matrix values, and located at the base of the phylogenetic trees, so it can be speculated that the C. sinensis may be the ancestral form in the genus Craspedacusta.

Construction of cytoplasmic molecular markers distinguishing Danio rerio from Gobiocypris rarus at high identity domains based on MP-PCR strategy and Sybr Green I detection

To distinguish the cytoplasm of Danio rerio from that of Gobiocypris rarus, we cloned G. rarus COXI and constructed cytoplasmic molecular markers at the high identity domains of COXI by mutated primer PCR (MP-PCR for short). Then Sybr Green I was used to detect the single amplicon. As a result, we succeeded in getting the cytoplasmic molecular markers, G.M COXI and Z.M COXI, by MP-PCR strategy. They were used to detect the sperm-derived mtDNA in the sexual hybrid embryos (D. rerio female x G. rarus male) before the sphere stage. In the present study, all results demonstrate that MP-PCR approach and Sybr Green I detection are feasible to construct the molecular markers to identify genes that shared high identity.

Developmental expression of CagMdkb during gibel carp embryogenesis

Midkine (Mdk) genes have been revealed to have different expression patterns in vertebrates and therefore, additional studies on Mdk expression patterns are required in more species. In this study, CagMdkb has been cloned and characterized from a SMART cDNA library of 10-somite stage embryos of Carassius auratus gibelio. Its full length cDNA is 1091 bp and encodes a sequence of 147 amino acids, which shows 97.3% identity to zebrafish Mdkb on the amino acid level. RT-PCR analysis reveals that CagMdkb is first transcribed in gastrula embryos and maintains a relatively stable expression level during subsequent embryogenesis. Western blot analysis reveals a 19 kDa maternal CagMdkb protein band and the zygotic CagMdkb protein is expressed from gastrula stage. At around 10 somite stage, the 19 kDa CagMdkb is processed to another protein band of about 17 kDa, which might be the secreted form with the 21-residue signal peptide removed. With immunofluorescence analysis, maternal CagMdkb protein was found to be localized in each blastamere cell of early embryos. The zygotic CagMdkb positive fluorescence signal was detected from a pair of large neurons at 18-somite stage. At the later stages, CagMdkb protein was also extended to numerous small neurons in the forebrain, midbrain and hindbrain, as well as to nerve fibers in the spinal cord. Co-localization with 3A10 antibody revealed CagMdkb immunoreactivity on developing Mauthner neurons, a member of reticulospinal neurons. In addition, ectopic expression of CagMdkb in early embryos of gibel carp and zebrafish suppressed head formation and CagMdkb function was found to depend on secretory activity. All these findings indicate that CagMdkb plays an important role in neural development during gibel carp embryogenesis and there is functional conservation of Mdkb in fish head formation.

Positive effects of continuous low nitrate levels on growth and photosynthesis of Alexandrium tamarense (Gonyaulacales, Dinophyceae)

The growth and photosynthesis of Alexandrium tamarense (Lebour) Balech in different nutrient conditions were investigated. Low nitrate level (0.0882 mmol/L) resulted in the highest average growth rate from day 0 to day 10 (4.58 x 10(2) cells mL(-1) d(-1)), but the lowest cell yield (5420 cells mL(-1)) in three nitrate level cultures. High nitrate-grown cells showed lower levels of chlorophyll a-specific and cell-specific light-saturated photosynthetic rate (P-m(chl a) and P-m(cell)), dark respiration rate (R-d(chl a) and R-d(cell)) and chlorophyll a-specific apparent photosynthetic efficiency (alpha(chl a)) than was seen for low nitrate-grown cells; whereas the cells became light saturated at higher irradiance at low nitrate condition. When cultures at low nitrate were supplemented with nitrate at 0.7938 mmol/L in late exponential growth phase, or with nitrate at 0.7938 mmol/L and phosphate at 0.072 mmol/L in stationary growth phase, the cell yield was drastically enhanced, a 7-9 times increase compared with non-supplemented control culture, achieving 43 540 cells mL(-1) and 52 300 cells mL(-1), respectively; however, supplementation with nitrate in the stationary growth phase or with nitrate and phosphate in the late exponential growth phase increased the cell yield by no more than 2 times. The results suggested that continuous low level of nitrate with sufficient supply of phosphate may facilitate the growth of A. tamarense.

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 characterization of Bacillus anthracis by multiplex PCR on DNA chip

Bacillus anthracis can be identified by detecting virulence factor genes located on two plasmids, pXO1 and pXO2. Combining multiplex PCR with arrayed anchored primer PCR and biotin-avidin alkaline phosphatase indicator system, we developed a qualitative DNA chip method for characterization of B. anthracis, and simultaneous confirmation of the species identity independent of plasmid contents. The assay amplifies pag gene (in pXO1), cap gene (in pXO2) and Ba813 gene (a B. anthracis specific chromosomal marker), and the results were indicated by an easy-to-read profile based on the color reaction of alkaline phosphatase. About 1 pg of specific DNA fragments on the chip wells could be detected after PCR. With the proposed method, the avirulent (pXO1(+)/2(-), pXO1(-)/2(+) and pXO1(-)/2(-)) strains of B. anthracis and distinguished 'anthrax-like' strains from other B. cereus group bacteria were unambiguously identified, while the genera other than Bacillus gave no positive signal. (C) 2004 Elsevier B.V. All rights reserved.

Development of a positive-negative selection procedure for gene targeting in fish cells

The gene targeting technique is a powerful tool for analyzing functions of cloned genes and for generating transgenic animals with site-directed integration of foreign genes. In order to develop this technique in fish, positive-negative selection (PNS) and homologous recombination vectors were constructed, and their expression was examined in fish cells. A vector (pNK) for PNS consists of the neomycin resistance gene (neo) as a positive selectable marker gene and the herpes simplex virus (HSV) thymidine kinase (tk) gene as a negative selectable marker gene. Positive selection with geneticin (G418) of epithelioma papulosum of carp (EPC) cells transfected with linearized pNK vector yielded 350 colonies, while double selection of transfected EPC cells with G418 and gancyclovir (Gc) resulted in nearly complete cell death, demonstrating that the PNS procedure is effective in fish cells. Homologous recombination vectors consist of the Xiphophorus melanoma receptor kinase (X mrk(Y)) gene as homologous sequence in addition to the neo and tk genes. Conditions for homologous recombination vector transfection and drug selection were established. After verification of the feasibility of expression of homologous recombination vectors in EPC cells, the first gene targeting experiments were attempted in the Xiphophorus melanoma cell line, PSM. Positive-negative selection of the targeting vector-transfectants led to a low enrichment in this particular cell line. The reasons for the low enrichment in PSM cells were discussed. (C) 2002 Elsevier Science B.V. All rights reserved.

Positive magnetoresistance in In0.53Ga0.47As/In0.52Al0.48As quantum well

Magnetotransport measurements have been carried out on In0.53Ga0.17As/In0.52Al0.48 As quantum wells in a temperature range between 1.5 and 77 K. We have observed a large positive magnetoresistance in the low magnetic field range, but saturating in high magnetic fields. The magnetoresistance results from two occupied subbands in the two-dimensional electron gas. With the intersubband scattering considered, we obtained the subband mobility by analyzing the positive magnetoresistance. It is found that the second subband mobility is larger than that of the first due to the existence of the intersubband scattering.

Effect of implantation of nitrogen into SIMOX buried oxide on its fixed positive charge density

In order to obtain greater radiation hardness for SIMOX (separation by implanted oxygen) materials, nitrogen was implanted into SIMOX BOX (buried oxide). However, it has been found by the C-V technique employed in this work that there is an obvious increase of the fixed positive charge density in the nitrogen-implanted BOX with a 150 out thickness and 4 x 10(15) cm(-2) nitrogen implantation dose, compared with that unimplanted with nitrogen. On the other hand, for the BOX layers with a 375 nm thickness and implanted with 2 x 10(15) and 3 x 10(15) cm(-2) nitrogen doses respectively, the increase of the fixed positive charge density induced by implanted nitrogen has not been observed. The post-implantation annealing conditions are identical for all the nitrogen-implanted samples. The increase in fixed positive charge density in the nitrogen-implanted 150 nm BOX is ascribed to the accumulation of implanted nitrogen near the BOX/Si interface due to the post-implantation annealing process according to SIMS results. In addition, it has also been found that the fixed positive charge density in initial BOX is very small. This means SIMOX BOX has a much lower oxide charge density than thermal SiO2 which contains a lot of oxide charges in most cases.

identity based threshold proxy signature

IEEE Computer Society

Large positive and negative lateral displacements from total internal reflection configuration with a weakly absorbing dielectric film

It is theoretically shown that the simultaneously large positive and negative lateral displacements will appear when the resonant condition is satisfied for a TE-polarized light beam reflected from the total internal reflection configuration with a weakly absorbing dielectric film. Appearance of the enhanced negative lateral displacement is relative to the incidence angle, absorption of the thin Elm and its thickness. If we select an appropriate weakly absorbing dielectric film and its thickness, the simultaneously enhanced positive and negative lateral displacements will appear at different resonant angles. These phenomena may lead to convenient measurements and interesting applications in optical devices.

Controllable negative and positive group delay in transmission through a single quantum well at finite magnetic fields

We investigate the controllable negative and positive group delay in transmission through a single quantum well at the finite longitudinal magnetic fields. It is shown that the magneto-coupling effect between the longitudinal motion component and the transverse Landau orbits plays an important role in the group delay. The group delay depends not only on the width of potential well and the incident energy, but also on the magnetic-field strengthen and the Landau quantum number. The results show that the group delay can be changed from positive to negative by the modulation of the magnetic field. These interesting phenomena may lead to the tunable quantum mechanical delay line. (c) 2007 Elsevier B.V. All rights reserved.