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1.
J Colloid Interface Sci ; 663: 601-608, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38428117

RESUMO

Li-rich layered oxides cathodes (LLOs) as the promising next-generation cathode materials can provide ultrahigh capacity and energy density due to their distinctive anionic redox chemistry. Unfortunately, severe interfacial side reactions, surface structural degradation and sluggish Li+ kinetics have resulted in low initial coulombic efficiency (ICE), capacity decay and poor rate performance, restricting their practical applications for high-energy-density lithium-ion batteries. Herein, Surface structure regulation strategy used as surface modified agent is proposed to activate the anionic redox chemistry via ammonium tungstate treatment. Experimental results showcase that dual coating layer spinel-like structure LiMn2O4 and Li2WO4 have been successfully constructed on the surface of LLOs. The surface spinel-like structure providing 3 D Li+ diffusion channels together with fast-ion conductive layer decrease the interfacial Li+ diffusion barrier and boost the fasting Li+ kinetics. In addition, the in-situ reconstruction layer can further alleviate the interfacial side reactions and reinforce the surface structural stability. As a result, the ICE of modified LLOs can be precisely increased from 74.71 % to 107.42 % with the adjustment of ammonium tungstate usage. Moreover, it delivers a high reversible capacity of 279.5 mAh/g at 0.1 C, as well as excellent rate capability with capacity of 147.2 mAh/g at 5 C. This work provides a significant reference for designing high-energy-density LLOs via surface structure regulation strategy.

2.
Small ; : e2401132, 2024 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-38552226

RESUMO

Li-rich layered oxides cathodes (LLOs) have prevailed as the promising high-energy-density cathode materials due to their distinctive anionic redox chemistry. However, uncontrollable anionic redox process usually leads to structural deterioration and electrochemical degradation. Herein, a Mo/Cl co-doping strategy is proposed to regulate the relative position of energy band for modulating the anionic redox chemistry and strengthening the structural stability of Co-free Li1.16Mn0.56Ni0.28O2 cathodes. The incorporation of Mo with high d state orbit and Cl with low electronegativity can narrow the band energy gap between bonding and antibonding bands via increasing the filled lower-Hubbard band (LHB) and decreasing the non-bonding O 2p energy bands, promoting the anionic redox reversibility. In addition, strong covalent Mo─O and Mn─Cl bonding further increases the covalency of Mn─O band to further stabilize the O2 n- species and enhance the reversible distortion of MnO6 octahedron. The strengthening electronic conductivity, together with the epitaxial structure Li2MoO4 facilitates the fast Li+ kinetics. As a result, the dual doping material exhibits enhanced anionic redox reversibility and suppressed oxygen release with increased cyclic stability and excellent rate performance. This strategy provides some guidance to design high-energy-density LLOs with desirable anionic redox reversibility and stable crystal structure via band structure engineering.

3.
Ying Yong Sheng Tai Xue Bao ; 34(8): 2267-2273, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37681391

RESUMO

As a vulnerable species identified by the International Union for Conservation of Nature (IUCN), Lepidochelys olivacea has attracted extensive attention in recent years. To examine its current distribution and that under future climate change scenarios, we compiled the occurrence data of L. olivacea. With eight predictor variables, including depth, offshore distance, mean primary productivity, minimum primary productivity, mean sea surface temperature, minimum sea surface temperature, mean sea surface salinity, and minimum sea surface salinity, we predicted its distribution in an ensemble species distribution model. The accuracy of the model was evaluated with the parameters of areas under curves (AUC) and true skill statistics (TSS). The results showed that the AUC and TSS values were 0.96 and 0.81, respectively, indicating a good predictive performance of the ensemble model. Sea surface temperature and salinity were the two most important variables determining the distribution of L. olivacea, with the suitable temperature ranging from 23 to 29 ℃ and salinity below 34. The current distribution range of L. olivacea was between 30° N-25° S. Under future climate scenarios, its distribution range would decrease, especially under the RCP85 scenario in the 2100s (with a 28% reduction in the suitable survival range). The results of model validation showed that it had high accuracy and could make accurate predictions of the distribution. This study would provide references for the development of more rational conservation measures and management strategies.


Assuntos
Mudança Climática , Salinidade , Temperatura
4.
Small ; 19(41): e2303539, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37287389

RESUMO

On account of high capacity and high voltage resulting from anionic redox, Li-rich layered oxides (LLOs) have become the most promising cathode candidate for the next-generation high-energy-density lithium-ion batteries (LIBs). Unfortunately, the participation of oxygen anion in charge compensation causes lattice oxygen evolution and accompanying structural degradation, voltage decay, capacity attenuation, low initial columbic efficiency, poor kinetics, and other problems. To resolve these challenges, a rational structural design strategy from surface to bulk by a facile pretreatment method for LLOs is provided to stabilize oxygen redox. On the surface, an integrated structure is constructed to suppress oxygen release, electrolyte attack, and consequent transition metals dissolution, accelerate lithium ions transport on the cathode-electrolyte interface, and alleviate the undesired phase transformation. While in the bulk, B doping into Li and Mn layer tetrahedron is introduced to increase the formation energy of O vacancy and decrease the lithium ions immigration barrier energy, bringing about the high stability of surrounding lattice oxygen and outstanding ions transport ability. Benefiting from the specific structure, the designed material with the enhanced structural integrity and stabilized anionic redox performs an excellent electrochemical performance and fast-charging property..

5.
Sci Total Environ ; 875: 162531, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36870512

RESUMO

The tissue distribution (liver, kidney, heart, lung, and muscle), source, and trophic transfer of 15 polycyclic aromatic hydrocarbons (PAHs) were studied on 14 stranded East Asian finless porpoises (Neophocaena asiaeorientalis sunameri), 14 spotted seals (Phoca largha), and 9 stranded minke whales (Balaenoptera acutorostrata) from Yellow Sea and Liaodong Bay. The PAHs levels ranged from below the limit of detection to 459.22 ng g-1 dry weight in the tissues of the three marine mammals, and light molecular weight PAHs were the primary pollutants. Although the PAHs levels were relatively higher in internal organs of the three marine mammals, generally no tissue-specific distribution of the PAHs congeners was found, either for gender-specific distribution of PAHs in the East Asian finless porpoises. However, species specific PAHs concentration distribution were obtained. The PAHs were mainly originated from petroleum and biomass combustion in the East Asian finless porpoises, while those for the spotted seals and minke whales were complex. Trophic level associated biomagnification was found for phenanthrene, fluoranthene, and pyrene in the minke whale. Benzo(b)fluoranthene exhibited a significant biodilution with increasing trophic levels in the spotted seals, but the total concentration of the PAHs showed a significant biomagnification with increasing trophic levels. Trophic level-associated biomagnification of acenaphthene, phenanthrene, anthracene, and ∑PAHs were found in the East Asian finless porpoise, while pyrene exhibited obvious biodilution with increasing trophic levels. Our current study filled knowledge gaps on tissue distribution and trophic transfer of the PAHs in the investigated three marine mammals.


Assuntos
Baleia Anã , Fenantrenos , Hidrocarbonetos Policíclicos Aromáticos , Toninhas , Focas Verdadeiras , Animais , Hidrocarbonetos Policíclicos Aromáticos/análise , Pirenos , Monitoramento Ambiental
6.
J Colloid Interface Sci ; 640: 373-382, 2023 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-36867934

RESUMO

Li-rich Mn-based layered oxides (LLOs) have emerged as one of the most promising cathode materials for the next-generation lithium-ion batteries (LIBs) because of their high energy density, high specific capacity, and environmental friendliness. These materials, however, have drawbacks such as capacity degradation, low initial coulombic efficiency (ICE), voltage decay, and poor rate performance due to irreversible oxygen release and structural deterioration during cycling. Herein, we present a facile method of triphenyl phosphate (TPP) surface treatment to create an integrated surface structure on LLOs that includes oxygen vacancies, Li3PO4, and carbon. When used for LIBs, the treated LLOs show an increased initial coulombic efficiency (ICE) of 83.6% and capacity retention of 84.2% at 1C after 200 cycles. It is suggested that the enhanced performance of the treated LLOs can be attributed to the synergetic functions of each component in the integrated surface, such as the oxygen vacancy and Li3PO4 being able to inhibit the evolution of oxygen and accelerate the transport of lithium ions, while the carbon layer can restrain undesirable interfacial side reactions and reduce the dissolution of transition metals. Furthermore, electrochemical impedance spectroscopy (EIS) and galvanostatic intermittent titration technique (GITT) prove an enhanced kinetic property of the treated LLOs cathode, and ex-situ X-ray diffractometer shows a suppressed structural transformation of TPP-treated LLOs during the battery reaction. This study provides an effective strategy for constructing an integrated surface structure on LLOs to achieve high-energy cathode materials in LIBs.

7.
J Colloid Interface Sci ; 615: 141-150, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35124502

RESUMO

Nowadays fast charging has become an important characteristic of lithium-ion batteries (LIBs), so is of great significance to study the fast charging of LIBs. However, previous research of fast charging has focused more on high energy density LIBs, due to the growing demand for electric vehicles. Herein, the fast-charging properties under ambient temperature and high temperature for (60 mAh LiCoO2/graphite batteries) micro-LIBs are firstly investigated. The electrochemical test results reveal that this kind of battery possesses 4C fast-charging capability. Further increase in charging rate will accelerate battery capacity decay without reducing charging time. Although high temperature increases the fast-charging capacity and shortens the fast-charging time to 10 min at 6C under 65 °C, increase of side reactions resulted from high temperature also exacerbates the performance of battery. Post-mortem analysis further demonstrates the structural changes of cathode and anode materials, residual lithium deposits, peeling of graphite and the incrassation of the solid electrolyte interphase (SEI), especially under high temperature, which lead to fast -charging performance degradation. This work reveals the possible causes of micro battery performance deterioration during fast charging under ambient and high temperature and provides some reference for designing micro-LIBs with fast -charging properties.

8.
J Colloid Interface Sci ; 606(Pt 2): 1729-1736, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34500171

RESUMO

During the production of micro lithium-ion batteries (LIBs), which are widely used in wireless headphones and other small portable devices, numerous factors can affect their quality, among which the content of water plays a crucial role. In this work, the influence of water in electrodes on the performances of micro LIBs is studied deeply. When the content of water increases, both the rate performance and the cycling performance of the batteries fade. The discharge capacity retention of the battery from high water content sample group H (group H) is 81.81% after 350 cycles at 2C, while that of the battery from low water content sample group L (group L) is 89.89% under the same condition. As for the rate performance, the discharge capacity of group H is only 58.66% of group L at 5C. To take a step further, it is mainly because an overgrowth of the solid electrolyte interphase film happen with the growth of water content. Accordingly, excess lithium ions are consumed and the porous structure of the anode is destroyed. Considering the results above, we believe that this work can offer a theory foundation to carry out the failure analysis of micro batteries.

9.
Microbiologyopen ; 10(2): e1190, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33970544

RESUMO

Age is known to affect the gut microbiota in various animals; however, this relationship is poorly understood in seabirds. We investigated the temporal succession of gut microbiota in captive chinstrap penguins of different ages using high-throughput sequencing. The gut microbiota exhibited a significant age succession pattern, reaching maturity in adults and then declining with increasing age. Only 15 amplicon sequence variants were shared among the gut microbiota in chinstrap penguins at all studied ages, and these contributed to most of the age-related variations in total gut microbiota. Co-occurrence networks found that these key bacteria belonged to the genera Acinetobacter, Clostridium sensu stricto, and Fusobacterium, and more species interactions were found within the same taxonomy. Functional prediction indicated that most of the metabolic functions were more abundant in the gut microbiota in adult chinstrap penguins, except for carbohydrate metabolism, which was significantly more abundant in older individuals.


Assuntos
Bactérias/classificação , Fezes/microbiologia , Microbioma Gastrointestinal , Metabolismo , Spheniscidae/microbiologia , Fatores Etários , Ração Animal , Animais , Bactérias/genética , DNA Bacteriano , DNA Ribossômico , Sequenciamento de Nucleotídeos em Larga Escala , Metagenoma , Interações Microbianas
10.
Braz. arch. biol. technol ; 64: e21210292, 2021. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1278439

RESUMO

Abstract NADPH-cytochromeP450 reductase (CPR) is one of the most important components of the cytochrome P450 enzyme system. In this study, a gene encoding CPR (named EsCPR) was isolated from Eriocheir sinensis using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. Analysis of the nucleotide sequence revealed a cDNA full-length of 3717 bp with an open reading frame of 2046 bp, a 5′-untranslated region of 42 bp, and a long 3′-untryganslated region of 1628bp, which encodes a protein of 681 amino acids with a predicted molecular weight of 30.7 kDa and an estimated pI of 4.82. The mature peptide shares amino acid of E. sinensis identity 82 % - 89 % to the CPR from Penaeus vannamei and Chionoecetes opilio. Tissues and developmental stage-dependent expression of EsCPR mRNA was investigated by real-time quantitative PCR. EsCPR mRNA was markedly expressed in the hepatopancreas and stomach. These results would provide valuable information for further study on the interactions between CPR and cytochrome P450 enzyme systems.


Assuntos
NADPH-Ferri-Hemoproteína Redutase , Clonagem de Organismos , Braquiúros , Expressão Gênica , Reação em Cadeia da Polimerase Via Transcriptase Reversa
11.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(5): 3648-9, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-26329595

RESUMO

In this study, the complete mitochondrial genome of Saxidomus purpuratus is determined, which is the first complete mitochondrial genome in the genus Saxidomus. The genome was of 19 637 bp in length, including 2 rRNAs, 22 tRNAs and 12 protein-coding genes with the order of ND3 and ND5 reversed. Maximum likelihood tree based on nucleotide sequences of 12 mitochondrial PCGs was constructed, in which S. purpuratus was clustered with 3 Meretrix species. The results are expected to provide useful data for species identification and further studies of the genus Saxidomus.


Assuntos
Bivalves/genética , Genoma Mitocondrial , Animais , Genes de RNAr , Tamanho do Genoma , NADH Desidrogenase/genética , RNA de Transferência/genética , Sequenciamento Completo do Genoma
12.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(5): 3569-70, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-26260176

RESUMO

In this study, we first determined and described the complete mitogenome sequence of Parastichopus californicus, which was 16 727 bp in length. The mitochondrial genome had the canonical mitochondrial gene content, including 13 protein-coding genes, 2 rRNA genes and 22 tRNA genes. The overall base composition of the heavy-strand was 31.5% A, 18 % G, 20.6% C and 29.9% T, with a high A + T content of 61.4%. ML phylogenetic tree indicated that P. californicus and P. nigripunctatus were clustered in one branch belonging to the genus Parastichopus. This conclusion was identical to the former result by the methods of morphological taxonomy.


Assuntos
Genoma Mitocondrial , Pepinos-do-Mar/genética , Animais , Composição de Bases , DNA Mitocondrial/genética , DNA Mitocondrial/isolamento & purificação , Genes Mitocondriais , Filogenia , RNA de Transferência/genética , Sequenciamento Completo do Genoma
13.
Mol Biol Rep ; 40(12): 6987-95, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24178342

RESUMO

The glutathione S-transferase (GST) fusion protein system is widely used for high-level expression and efficient purification of recombinant proteins from bacteria. The goal of this study was to clone, efficiently express and purify the ecdysteroid-regulated protein (ERP) in the form of a GST fusion protein. The mature peptide-coding cDNA fragment was extracted from Chinese mitten crap (Eriocheir sinensis), and then after using PCR to obtain the open reading frame, a recombinant plasmid designated pGEX-4T-1_ERP was successfully generated and showed to efficiently express the ERP fusion protein as determined by SDS-PAGE. The resulting expressed protein was successfully purified by a combination of affinity and conventional chromatographic methods. After purification, the recombinant protein showed the expected size of 41 kDa on SDS-PAGE gels which was further confirmed by mass spectrometry and western blotting. Purification of recombinant protein was achieved by fast protein liquid chromatography. About 2.4 mg/l recombinant protein with purity more than 80 % was obtained.


Assuntos
Proteínas de Artrópodes/isolamento & purificação , Proteínas de Artrópodes/metabolismo , Braquiúros/metabolismo , Ecdisteroides/metabolismo , Escherichia coli/metabolismo , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Sequência de Bases , China , Cromatografia Líquida , Clonagem Molecular , DNA/metabolismo , DNA Complementar/genética , Vetores Genéticos/metabolismo , Espectrometria de Massas , Dados de Sequência Molecular , Filogenia , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA
14.
Mol Biol Rep ; 40(8): 4769-79, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23653004

RESUMO

Tumor necrosis factor receptor-associated factor 6 (TRAF6) is a key adaptor molecule for the tumor necrosis factor superfamily and Toll-like/interleukin-1 receptor superfamily. It plays an important role in innate and adaptive immunity. The TRAF6 of Japanese scallop Mizuhopecten yessoensis (designated as MyTRAF6) was identified and characterized in this study. The full-length cDNA of MyTRAF6 was 2,407 bp, which consisted of 239-bp 5'-terminal untranslated region, 1,974-bp open reading frame encoding a polypeptide of 657 amino acids, 194-bp of 3'-terminal untranslated region followed by a canonical polyadenylation signal sequence AATAAA and a poly (A) tail. The predicted amino acid sequence of MyTRAF6 contained the characteristic motifs of TRAF proteins, including a Zinc finger of RING-type, two Zinc fingers of TRAF-type, and a MATH (meprin and TRAF homology) domain. It had an overall identity of 43-96% with those of other TRAF6s, the highest identity (96%) with Chlamys farreri TRAF6, and the least identity (43%) with Meleagris gallopavo TRAF6. Phylogenetic analysis classified MyTRAF6 as a true TRAF6 ortholog. In addition, the promoter of MyTRAF6 was also identified by genome walking. It contained several potential transcription factor-binding sites and three single nucleotide polymorphisms. qRT-PCR analysis revealed that MyTRAF6 was highly expressed in hemocytes of adult M. yessoensis. MyTRAF6 transcript level in the hemocytes reached a maximum 6 h after Vibrio anguilarum challenge. The results indicated that MyTRAF6 may fulfill an important function during M. yessoensis bacterial infection. It could be a key effector molecule involved in the innate defense of molluscs.


Assuntos
Bivalves/genética , Imunidade Inata/genética , Filogenia , Conformação Proteica , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Sítios de Ligação/genética , Bivalves/imunologia , Bivalves/microbiologia , Clonagem Molecular , DNA Complementar/genética , Hemócitos/metabolismo , Japão , Dados de Sequência Molecular , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Homologia de Sequência , Vibrio/imunologia
15.
Artigo em Inglês | MEDLINE | ID: mdl-23396151

RESUMO

Spotted seal (Phoca largha) is categorized as a critically endangered species in China. The aim of this study was to investigate spotted seal transcriptome by the approach of Illumina paired-end sequencing technology. We obtained a total of 52,146,394 reads for the mixed tissues of liver and spleen from the spotted seal. The de novo assemblies yielded 354,014 contigs and 178,466 unigenes. In the transcriptome, 193 unigenes were assigned to defense mechanisms. Three unigenes encoded MHC class I and 17 unigenes encoded MHC class II. In addition, bioinformatics analysis revealed a total of 4425 simple sequence repeats (SSRs). Fifty SSRs were randomly selected to validate amplification and determine the degree of polymorphism in the genomic DNA pools. Thirty-five primer pairs successfully amplified the expected DNA fragments and detected significant polymorphism among 28 spotted seal individuals. These results would contribute to the understanding of the genetic makeup of spotted seal transcriptome and provide useful information for functional genomic research in this species.


Assuntos
Phoca/genética , Transcriptoma , Animais , DNA Complementar/análise , DNA Complementar/genética , DNA Complementar/metabolismo , Bases de Dados Genéticas , Etiquetas de Sequências Expressas , Marcadores Genéticos/genética , Fígado/química , Fígado/metabolismo , Masculino , Repetições de Microssatélites , Família Multigênica , Phoca/metabolismo , Polimorfismo Genético , RNA/genética , RNA/isolamento & purificação , Reprodutibilidade dos Testes , Análise de Sequência de DNA/métodos , Baço/química , Baço/metabolismo
16.
Int J Mol Sci ; 13(11): 14172-202, 2012 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-23203058

RESUMO

Catfish is one of the most important aquaculture species in America (as well as in Asia and Africa). In recent years, the production of catfish has suffered massive financial losses due to pathogen spread and breakouts. Innate immunity plays a crucial role in increasing resistance to pathogenic organisms and has generated increasing interest in the past few years. This review summarizes the current understanding of innate immune-related genes in catfish, including pattern recognition receptors, antimicrobial peptides, complements, lectins, cytokines, transferrin and gene expression profiling using microarrays and next generation sequencing technologies. This review will benefit the understanding of innate immune system in catfish and further efforts in studying the innate immune-related genes in fish.


Assuntos
Peixes-Gato/genética , Peixes-Gato/imunologia , Imunidade Inata/genética , Animais , Perfilação da Expressão Gênica , Regulação da Expressão Gênica
17.
Mol Biol Rep ; 39(12): 10713-22, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23053968

RESUMO

Hard clam (Meretrix meretrix) is an economically important bivalve in China. In the present study, a gene coding for an intracellular Cu/Zn-SOD was cloned and characterized from hard clam. The full-length cDNA of this Cu/Zn-SOD (designated as Mm-icCuZn-SOD) consisted of 1,383 bp, with a 462-bp of open reading frame (ORF) encoding 153 amino acids. Several highly conserved motifs, including the Cu/Zn binding sites [H(46), H(48), H(63), and H(119) for Cu binding; H(63), H(71), H(80), and D(83) for Zn binding], an intracellular disulfide bond and two Cu/Zn-SOD signatures were identified in Mm-icCu/Zn-SOD. The deduced amino acid sequence of Mm-icCu/Zn-SOD has a high degree of homology with the Cu/Zn-dependent SODs from other species, indicating that Mm-icCu/Zn-SOD should be a member of the intracellular Cu/Zn-dependent SOD family. Real-time PCR analysis showed that the highest level of Mm-icCu/Zn-SOD expression was in the hepatopancreas, while the lowest level occurred in the hemocytes. Hard clam challenged with Vibrio anguillarum showed a time-dependent increase in Mm-icCu/Zn-SOD expression that reached a maximum level after 6 h. Mm-icCu/Zn-SOD purified as a recombinant protein expressed in E. coli retained a high level of biological activity, 83 % after 10 min incubation at 10-50 °C, and more than 87 % after incubation in buffers with pH values between 2.2 and 10.2. These results indicated that Mm-icCu/Zn-SOD may play an important role in the innate immune system of hard clam.


Assuntos
Bivalves/enzimologia , Bivalves/genética , DNA Complementar/genética , Regulação Enzimológica da Expressão Gênica , Espaço Intracelular/enzimologia , Superóxido Dismutase/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Clonagem Molecular , Estabilidade Enzimática , Perfilação da Expressão Gênica , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Superóxido Dismutase/química , Superóxido Dismutase/isolamento & purificação , Temperatura , Fatores de Tempo
18.
Artigo em Inglês | MEDLINE | ID: mdl-22677091

RESUMO

Next-generation sequencing provides a powerful new approach for developing functional genomic tools for non-model species. This study aims to investigate the spotted seal (Phoca largha) transcriptome by the approach of Illumina paired-end sequencing technology. We obtained a total of 52,146,394 reads for the mixed tissues of liver and spleen from spotted seal. The de novo assemblies yielded 354,014 contigs and 178,466 unigenes. In addition, bioinformatics analysis revealed a total of 4425 simple sequence repeats (SSRs). Fifty SSRs were randomly selected to validate amplification and determine the degree of polymorphism in the genomic DNA pools. Thirty-five primer pairs successfully amplified expected DNA fragments and detected significant polymorphism among 28 spotted seal individuals. These results contribute to the understanding of the genetic makeup of spotted seal transcriptome and provide useful information for functional genomic research in this species.


Assuntos
Repetições de Microssatélites/genética , Phoca/genética , Análise de Sequência de DNA/métodos , Transcriptoma/genética , Animais , Sequência de Bases , Análise por Conglomerados , Marcadores Genéticos , Complexo Principal de Histocompatibilidade/genética , Anotação de Sequência Molecular , Motivos de Nucleotídeos/genética , Nucleotídeos/genética , Polimorfismo Genético
19.
Comp Biochem Physiol B Biochem Mol Biol ; 162(1-3): 34-43, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22366552

RESUMO

Lysozyme is an important component of the immune response against bacteria that is characterized by its ability to break down bacterial cell-walls. We constructed a high-quality cDNA library from mantle tissue of adult Japanese scallop (Mizuhopecten yessoensis). The EST which is high homology with g-type lysozyme genes of other species was found in the cDNA library. In the present study, the complete express sequence of g-type lysozyme genes from Japanese scallop (designated as MyLysoG) was directly obtained by PCR. The complete sequence of MyLysoG cDNA consisted of a 5' untranslated region (UTR) of 25 bp, an open reading frame (ORF) of 606 bp, and a 3' UTR of 100 bp with one polyadenylation signal (AATAAA). The deduced amino acids of MyLysoG were 201 amino acids with a putative signal peptide of 18 amino acid residues. It shared the sequence similarity and the common structure features with the g-type lysozyme from other species. Quantitative reverse trancriptase real-time PCR (qRT-PCR) assay demonstrated that mRNA transcripts of g-type lysozyme could be detected in various tissues of unchallenged scallop, and the highest expression of MyLysoG was detected in hepatopancreas tissue. The temporal expression of MyLysoG in hemolymph after Vibrio anguillarum challenge was up-regulated and reached the maximum level at 3h post stimulation, and then dropped back to the original level even lower than the control group. Furthermore, a 978 bp of 5'-flanking sequence of MyLysoG was identified by genome walking, and several potential transcription factor binding sites (TFBS) were detected in the putative promoter region. One part of the MyLysoG promoter region contains nine sites of SNPs and three sites of insert-deletion (indel) polymorphisms, and these mutations were found organize into two haplotypes. The two haplotypes were associated with different TFBS. The haplotypes could be selected to analyze the transcriptional-level control of scallop g-type lysozyme gene and the scallop immune system.


Assuntos
Clonagem Molecular , Muramidase/genética , Pectinidae/enzimologia , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Sequência de Aminoácidos , Animais , DNA Complementar/química , DNA Complementar/genética , DNA Complementar/metabolismo , Dados de Sequência Molecular , Pectinidae/genética , Pectinidae/metabolismo , Filogenia , Homologia de Sequência de Aminoácidos
20.
Mol Biol Rep ; 39(2): 865-71, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21567197

RESUMO

LPS-induced TNF-α (LITAF) is a novel transcriptional factor that mediates the expression of inflammatory cytokines in LPS-induced processes. In the present study, the full-length cDNA encoding LITAF (designated as Mm-LITAF) was identified from Asiatic hard clam, Meretrix meretrix, by expressed sequence tag and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Mm-LITAF was 1653 bp, consisting of a 5' untranslated region (UTR) of 91 bp, a 3'UTR of 1166 bp with one cytokine RNA instability motif (ATTTA) and one polyadenylation signal (AATAAA), and an open reading frame (ORF) of 396 bp encoding a polypeptide of 131 amino acids with a theoretical isoelectric point of 7.49, and predicted molecular weight of 14.47 kDa. The deduced amino acid of Mm-LITAF shared 29-63% similarity with the LITAFs from other species, indicating that Mm-LITAF should be a member of the LITAF family. Two highly conserved CXXC motifs forming a compact Zn(2+)-binding structure were also identified in Mm-LITAF. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of Mm-LITAF mRNA in different tissues, and the temporal expression of Mm-LITAF in clams challenged with Vibrio anguillarum. The mRNA transcript of Mm-LITAF could be detected in all the examined tissues with the highest expression level in the gill. Mm-LITAF expression was up-regulated significantly at 16 h in the gill and at 8 h in haemocytes after bacterial challenge, respectively. These results suggest that the Mm-LITAF is a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of hard clam.


Assuntos
Bivalves/genética , Filogenia , RNA Mensageiro/genética , Fatores de Transcrição/genética , Fator de Necrose Tumoral alfa/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bivalves/microbiologia , Clonagem Molecular , Análise por Conglomerados , Primers do DNA/genética , DNA Complementar/genética , Etiquetas de Sequências Expressas , Componentes do Gene , Brânquias/metabolismo , Hemócitos/metabolismo , Lipopolissacarídeos , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência , Análise de Sequência de DNA , Fatores de Transcrição/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
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