952 resultados para cross-species amplification
Resumo:
Broadband infrared luminescence is observed in various Bi-doped oxide glasses prepared by conventional melting-quenching technique. The absorption spectrum of the Bi-doped germanium oxide glass consists of five broad peaks at below 370, 500, 700, 800 and 1000 nm. The fluorescence spectrum exhibits a broad peak at about 1300 nm with full width at half maximum (FWHM) of more than 300 nm when excited by an 808 nm laser diode. The fluorescence lifetime at room temperature decreases with increasing Bi2O3 concentration. Influence of the glass composition and melting atmosphere on the fluorescence lifetime and luminescent intensity is investigated. The mechanism of the broadband infrared luminescence is suggested. The product of stimulated emission cross-section and lifetime of the Bi-doped aluminophosphate glass is about 5.0 X 10(-24) cm(2) s. The glasses might be promising for applications in broadband optical fiber amplifiers and tunable lasers. (c) 2007 Elsevier B.V. All rights reserved.
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The variability in the supply of pink shrimp (Farfantepenaeus duorarum) postlarvae and the transport mechanisms of planktonic stages were investigated with field data and simulations of transport. Postlarvae entering the nursery grounds of Florida Bay were collected for three consecutive years at channels that connect the Bay with the Gulf of Mexico, and in channels of the Middle Florida Keys that connect the southeastern margin of the Bay with the Atlantic Ocean. The influx of postlarvae in the Middle Florida Keys was low in magnitude and varied seasonally and among years. In contrast, the greater postlarval influx occurred at the northwestern border of the Bay, where there was a strong seasonal pattern with peaks in influx from July through September each year. Planktonic stages need to travel up to 150 km eastward between spawning grounds (northeast of Dry Tortugas) and nursery grounds (western Florida Bay) in about 30 days, the estimated time of planktonic development for this species. A Lagrangian trajectory model was developed to estimate the drift of planktonic stages across the SW Florida shelf. The model simulated the maximal distance traveled by planktonic stages under various assumptions of behavior. Simulation results indicated that larvae traveling with the instantaneous current and exhibiting a diel behavior travel up to 65 km and 75% of the larvae travel only 30 km. However, the eastward distance traveled increased substantially when a larval response to tides was added to the behavioral variable (distance increased to 200 km and 85% of larvae traveled 150 km). The question is, when during larval development, and where on the shallow SW Florida shelf, does the tidal response become incorporated into the behavior of pink shrimp.
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This study has developed an improved subjective approach of classification in conjunction with Step wise DFA analysis to discriminate Chinese sturgeon signals from other targets. The results showed that all together 25 Chinese sturgeon echo-signals were detected in the spawning ground of Gezhouba Dam during the last 3 years, and the identification accuracy reached 90.9%. In Stepwise DFA, 24 out of 67 variables were applied in discrimination and identification. PCA combined with DFA was then used to ensure the significance of the 24 variables and detailed the identification pattern. The results indicated that we can discriminate Chinese sturgeon from other fish species and noise using certain descriptors such as the behaviour variables, echo characteristics and acoustic cross-section characteristics. However, identification of Chinese sturgeon from sediments is more difficult and needs a total of 24 variables. This is due to the limited knowledge about the acoustic-scattering properties of the substrate regions. Based on identified Chinese sturgeon individuals, 18 individuals were distributed in the region between the site of Gezhouba Dam and Miaozui reach, with a surface area of about 3.4 km(2). Seven individuals were distributed in the region between Miaozui and Yanshouba reach, with a surface area of about 13 km(2).
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Artificial interspecific hybrids between large scale loach P. dabryanus and tetraploid pond loach M. anguillicaudatus (Cobitidae, Cypriniformes) are viable. To detect the occurrence of possible natural hybridization, genetic analyses by using microsatellite markers were performed for natural populations of large scale loach and pond loach, the reciprocal laboratory hybrids, and "supposed hybrids" with ambiguous morphology. The fertility of the artificial hybrids was also tested. At one diagnostic microsatellite (Mac50), one out of 20 "supposed hybrids" was identified to be F-1 hybrid between the two loach species because it had the same genotype as that of the laboratory hybrids. The triploid hybrids between the two species were confirmed to be female-sterile. The results show that rare hybridization has occurred between diploid large scale loach and tetraploid pond loach in nature although it may have little effect in genetic introgression. This study is helpful for fish conservation and encourages further investigation on natural hybridization and introgression of loaches.
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Background: Short and long interspersed elements (SINEs and LINEs, respectively), two types of retroposons, are active in shaping the architecture of genomes and powerful tools for studies of phylogeny and population biology. Here we developed special protocol to apply biotin-streptavidin bead system into isolation of interspersed repeated sequences rapidly and efficiently, in which SINEs and LINEs were captured directly from digested genomic DNA by hybridization to bead-probe complex in solution instead of traditional strategy including genomic library construction and screening. Results: A new couple of SINEs and LINEs that shared an almost identical 3'tail was isolated and characterized in silver carp and bighead carp of two closely related species. These SINEs (34 members), designated HAmo SINE family, were little divergent in sequence and flanked by obvious TSD indicated that HAmo SINE was very young family. The copy numbers of this family was estimated to 2 x 10(5) and 1.7 x 10(5) per haploid genome by Real-Time qPCR, respectively. The LINEs, identified as the homologs of LINE2 in other fishes, had a conserved primary sequence and secondary structures of the 3'tail region that was almost identical to that of HAmo SINE. These evidences suggest that HAmo SINEs are active and amplified recently utilizing the enzymatic machinery for retroposition of HAmoL2 through the recognition of higher-order structures of the conserved 42-tail region. We analyzed the possible structures of HAmo SINE that lead to successful amplification in genome and then deduced that HAmo SINE, SmaI SINE and FokI SINE that were similar in sequence each other, were probably generated independently and created by LINE family within the same lineage of a LINE phylogeny in the genomes of different hosts. Conclusion: The presented results show the advantage of the novel method for retroposons isolation and a pair of young SINE family and its partner LINE family in two carp fishes, which strengthened the hypotheses containing the slippage model for initiation of reverse transcription, retropositional parasitism of SINEs on LINEs, the formation of the stem loop structure in 3'tail region of some SINEs and LINEs and the mechanism of template switching in generating new SINE family.
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Triplophysa lixianensis, a new nemacheiline loach species, is described from the Min Jiang of the upper Yangtze River drainage in Sichuan Province, South China. It can be separated from all other species of Triplophysa by having a unique combination of the following characters: posterior chamber of gas bladder greatly reduced or absent; caudal peduncle columnar with a roughly round cross- section at its beginning; anterior edge of lower jaw completely exposed or uncovered by lower lip; intestine short, forming a zigzag loop below stomach; dorsal- fin origin closer to caudal- fin base than to snout tip; pelvic fin inserted anterior to dorsal- fin origin; snout length 50.6 - 57.5 % of head length; eye diameter 12.3 15.4 % of head length; caudal peduncle length 25.1 - 27.1 % of standard length; anal fin with five branched rays; lower lip greatly furrowed with two thick lateral lobes; and body smooth or scaleless.
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In previous studies of nuclear transplantation, most cloned animals were obtained by intraspecies nuclear transfer and are phenotypically identical to their nuclear donors; furthermore, there was no further report on successful fish cloning since the report of cloned zebrafish. Here we report the production of seven cross-genus cloned fish by transferring nuclei from transgenic common carp into enucleated eggs of goldfish. Nuclear genomes of the cloned fish were exclusively derived from the nuclear donor species, common carp, whereas the mitochondrial DNA from the donor carp gradually disappeared during the development of nuclear transfer (NT) embryos. The somite development process and somite number of nuclear transplants were consistent with the recipient species, goldfish, rather than the nuclear donor species, common carp. This resulted in a long-lasting effect on the vertebral numbers of the cloned fish, which belonged to the range of goldfish. These demonstrate that fish egg cytoplasm not only can support the development driven by transplanted nuclei from a distantly related species at the genus scale but also can modulate development of the nuclear transplants.
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Phylogenetic relationships within Metapenaeopsis remain largely unknown. The modern revision of the genus suggests that the shape of the petasma, followed by the presence of a stidulating organ, are the most important distinguishing taxonomic features. In the present study, phylogenetic relationships were studied among seven Metapenaeopsis species from the Indo-West Pacific based on partial sequences of mitochondrial 16S rRNA and cytochrome c oxidase I (COI) genes. Mean sequence divergence was 6.4% for 16S and 15.8% for COI. A strikingly large nucleotide distance (10.0% for 16S and 16.9% for COI) was recorded between M. commensalis, the only Indo-West Pacific species with a one-valved petasma, and the other species with a two-valved petasma. Phylogenetic analyses using neighbor-joining, maximum parsimony, and maximum likelihood generated mostly identical tree topologies in which M. commensalis is distantly related to the other species. Two clades were resolved for the remaining species, one with and the other without a stridulating organ, supporting the main groupings of the recent taxonomic revision. Results of the present study also indicate that the deep-water forms represent a relatively recent radiation in Metapenaeopsis.
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A method of loop-mediated isothermal amplification (LAMP) was employed to develop a rapid and simple detection system for porcine circovirus type 2 (PCV2). The amplification could be finished in 60 min under isothermal condition at 64 degrees C by employing a set of four primers targeting the cap gene of PCV2. The LAMP assay showed higher sensitivity than the conventional PCR, with a detection limit of five copies per tube of purified PCV2 genomic DNA. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 1 (PCV1), porcine parvovirus (PPV), porcine pseudorabies virus (PRV) and porcine reproductive and respiratory syndrome virus (PRRSV). The detection rate of PCV2 LAMP for 86 clinical samples was 96.5% and appeared greater than that of the PCR method. The LAMP assay reported can provide a rapid yet simple test of PCV2 suitable for laboratory diagnosis and pen-side detection due to ease of operation and the requirement of only a regular water bath or heat block for the reaction. (c) 2008 Elsevier B.V. All rights reserved.
Resumo:
Antibody was covalently immobilized by amine coupling method to gold surfaces modified with a self-assembled monolayer of thioctic acid. The electrochemical measurements of cyclic voltammetry and impedance spectroscopy showed that the hexacyanoferrate redox reactions on the gold surface were blocked due to the procedures of self-assembly of thioctic acid and antibody immobilization. The binding of a specific antigen to antibody recognition layer could be detected by measurements of the impedance change. A new amplification strategy was introduced for improving the sensitivity of impedance measurements using biotin labeled protein- streptavidin network complex. This amplification strategy is based on the construction of a molecular complex between streptavidin and biotin labeled protein. This complex can be formed in a cross-linking network of molecules so that the amplification of response signal will be realized due to the big molecular size of complex. The results show that this amplification strategy causes dramatic improvement of the detection sensitivity of hIgG and has good correlation for detection of hIgG in the range of 2-10 mug/ml. (C) 2001 Elsevier Science B.V. All rights reserved.
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The effects of radiation on chlorinated polyisobutylene were studied. It was found that polyisobutylene, which is generally considered to be a typical radiation-degradative polymer, could be crosslinked by gamma-ray irradiation after chlorine had been introduced into its macromolecules. Carbon-chlorine bonds act both as radiation-sensitive groups and reaction species in the radiation process of chlorinated polyisobutylene.
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General equations of the electrocatalytic reaction at an ultramicroelectrode modified with redox species have been described according to the Andrieux Saveant model. The electrocatalytic kinetic process has been discussed for the whole set of cases, ie (R), (R + S), (SR) (SR + E), (E), (R + E), (ER), (S), (ER + S) and (S + E) limiting situations. The effect of gamma on the catalytic steady state current shows that the higher the value of gamma, the lower the catalytic current. The kinetic process shifts rapidly from R to E with increasing values of gamma. It is favorable for catalysis only when gamma is very low. Therefore, the redox species modified ultramicroelectrode with thin film is utilized for electrocatalysis, and the larger the radius of ultramicroelectrode, the higher the catalytic efficiency.
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The current equation of the electrocatalytic reaction at a microdisk electrode modified with redox species has been described and verified experimentally. There exists a linear relationship between plateau limiting current and the radius of the microdisk electrode for a catalytic process. The influence of the dimensions of the microdisk electrode on catalytic efficiency is discussed. The polyvinylferrocene (PVFc)-modified microdisk electrode prepared by the coating method was taken as a typical example, on which the electrocatalytic oxidation of ascorbic acid could be studied. The catalytic reaction rate constants were determined as an average value of 1.5 X 10(-7) cm3/mol s by this method, and are consistent with those obtained at a conventional electrode.
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Karyotype and chromosomal location of the major ribosomal RNA genes (rDNA) were studied using fluorescence in situ hybridization (FISH) in five species of Crassostrea: three Asian-Pacific species (C. gigas, C. plicatula, and C. ariakensis) and two Atlantic species (C. virginica and C. rhizophorae). FISH probes were made by PCR amplification of the intergenic transcribed spacer between the 18S and 5.8S rRNA genes, and labeled with digoxigenin-11-dUTP. All five species had a haploid number of 10 chromosomes. The Atlantic species had 1-2 submetacentric chromosomes, while the three Pacific species had none. FISH with metaphase chromosomes detected a single telomeric locus for rDNA in all five species without any variation. In all three Pacific species, rDNA was located on the long arm of Chromosome 10 (10q)-the smallest chromosome. In the two Atlantic species, rDNA was located on the short arm of Chromosome 2 (2p)-the second longest chromosome. A review of other studies reveals the same distribution of NOR sites (putative rDNA loci) in three other species: on 10q in C. sikamea and C. angulata from the Pacific Ocean and on 2p in C. gasar from the western Atlantic. All data support the conclusion that differences in size and shape of the rDNA-bearing chromosome represent a major divide between Asian-Pacific and Atlantic species of Crassostrea. This finding suggests that chromosomal divergence can occur under seemingly conserved karyotypes and may play a role in reproductive isolation and speciation.
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A Gymnodinium-like species was studied with light microscopy (LM) and scanning electron microscopy (SEM). Also, the internal transcribed spacers (containing 5.8S rDNA) and large ribosomal subunit DNA (D1-D2) sequences were obtained by PCR amplification, and then sequenced to explore the relationships within our isolate, Gymnodinium and other Gymnodinium-like species, including Karenia, Gyrodinium, Karlodinium and Symbiodinium. The LM observation showed that the species was characterized by moving in a levorotatory direction, visible hypocone, epicone and transverse groove, all of which are typical for Gymnodinium. In addition, two flagella could be found under SEM. The phylogenetic analysis revealed that the isolate grouped with Symbiodium, rather than other relevant dinoflagellates. All results showed our isolate belongs to Symbiodium. The strain was isolated from a red tide water sample, denoting that Symbiodium may be causative species for algal bloom.