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Design and Expression of a Synthetic phyC Gene Encoding the Neutral Phytase in Pichia pastoris

Acta Biochimica et Biophysica Sinica 2006年第11期38卷 803-811页

作者： Li-Kou ZOU~(1,3) Hong-Ning WANG~(2,1*) Xin PAN~4 Tao XIE~1 Qi WU~5 Zi-Wen XIE~1 Wan-Rong ZHOU~1 1 Laboratory of Veterinary and Biotechnology,sichuan Agricultural university,Ya'an 625014,china 2 sichuan university bioengineering research center for animal disease prevention and control,chengdu 610065,china 3 Faculty of Environment and Resource of Dujiangyan Campus,sichuan Agricultural university,Dujiangyan 611830,china 4 College of Earth Science,Chengd university of Technology,chengdu 610059,china 5 College of Life Science,sichuan Agricultural university,Ya'an 625014,china Laboratory of Veterinary and Biotechnology Sichuan Agricultural University Ya'an 625014 China

The 1074-bp phyCs gene （optimized phyC gene） encoding neutral phytase was designed andsynthesized according to the methylotrophic yeast Pichia pastoris codon usage bias without altering theprotein *** expression vector,pP9K-phyCs,was linearized and transformed in *** of total extracellular phytase activity was 17.6 U/ml induced in Buffered Methanol-complex Medium（BMMY） and 18.5 U/ml in Wheat Bran Extract Induction （WBEI） medium at the flask scale,respectively,improving over 90 folds compared with the wild-type *** enzyme showed temperature optimumof 70℃ and pH optimum of *** enzyme activity retained 97% of the relative activity afterincubation at 80℃ for 5 *** of the heavy glycosylation the expressed phytase had a molecularsize of approximately 51 *** deglycosylation by endoglycosylase H （EndoH_f）,the enzyme had anapparent molecular size of 42 *** property and thermostability was affected by the glycosylation.

关键词： design expression synthetic phyC gene neutral phytase Pichia pastoris

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Characterization of codon usage bias in the dUTPase gene of duck enteritis virus

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Progress in Natural Science:Materials International 2008年第9期18卷 1069-1076页

作者： Lichan Zhao a, Anchun Cheng a,b,, Mingshu Wang a,b,, Guiping Yuan c, Mingsheng Caia a Avian disease research center, College of Veterinary Medicine of sichuan Agricultural university, Xinkang Road 46, Ya’an, sichuan 625014, china b Key Laboratory of animal disease and Human Health of sichuan Province, Ya’an 625014, china c Analytical & Testing center, sichuan university, chengdu 610064, china Avian disease research center College of Veterinary Medicine of Sichuan Agricultural University Xinkang Road 46 Ya’an Sichuan 625014 China Key Laboratory of animal disease and Human Health of sichuan Province Ya’an 625014 China Analytical & Testing center Sichuan University Chengdu 610064 China

A comparative analysis of the codon usage bias in the newly discovered dUTPase gene (Assigned Accession No.:DQ486149) of the duck enteritis virus (DEV) and the dUTPase gene of 32 reference herpesviruses was *** results indicated that the DEV dUT-Pase gene encodes a protein of 477 amino acids, which includes five conserved motifs with a 3-1-2-4-5 *** codon adap-tation index (CAI), efiective number of codons (ENC), and GC3S values indicated synonymous codon usage bias in the dUTPase gene of herpesviruses, and this synonymous bias was correlated with host *** codon usage patterns of the DEV dUTPase gene were phylogenetically conserved and similar to that of the dUTPase genes of the avian *** codon usage in each micro-organism was difierent, there were no strain-specific difierences among ***-one codons in the predicted polypeptide, with a strong bias towards A and T at the third codon position, were *** of the codon usage in the dUTPase gene of difierent organisms revealed that there were 19 codons showing distinct codon usage difierences between the DEV and Escherichia coli dUTPase genes;16 between the DEV and yeast dUTPase genes;and 15 between the DEV and human dUTPase *** of variance (ANOVA) showed significant difierences between the DEV and yeast dUTPase genes (r = 0.536, P < 0.01).The extent of codon usage bias in the DEV dUTPase gene was highly correlated with the gene expression level, therefore the results may provide useful information for gene classification and functional studies.

关键词： DEV dUTPase Codon usage bias EMBOSS

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Dynamic changes of apoptosis in duck embryo fibroblasts induced by new type Gosling viral enteritis virus

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Progress in Natural Science:Materials International 2008年第2期18卷 239-244页

作者： Shun Chen a,Anchun Cheng a,b,,Mingshu Wang a,b,Xiaoyue Chen a a Avian disease research center,College of Veterinary Medicine of sichuan Agricultural university,Yaan 625014,china b Key Laboratory of animal disease and Human Health of sichuan Province,Yaan 625014,china Avian disease research center College of Veterinary Medicine of Sichuan Agricultural University Yaan 625014 China Key Laboratory of animal disease and Human Health of sichuan Province Yaan 625014 China

The monolayer duck embryo fibroblast (DEF) cells were experimentally infected with new type Gosling viral enteritis virus (NGVEV) and the dynamic changes of apoptosis were detected at different time points after NGVEV infection by transmission electron microscopy (TEM),DNA agarose gel electrophoresis and Annexin V-FITC/PI stained fluorescence-activated cell sorter (FACS). The result shows that NGVEV can induce infected cells undergoing apoptosis and changing regularly. A series of characteristic apoptotic morphological changes including shrinkage of the cells,chromatin condensation and margination,as well as formation of apoptotic bodies,were observed by TEM. The typical ladder pattern of DNA fragmentation was demonstrated by agarose gel electrophoresis. And using flow cytometry analysis of Annexin V-FITC/PI staining,the dead,viable,apoptotic and necrotic cells could be analyzed quantitatively.

关键词： New type Gosling viral enteritis virus (NGVEV) Duck embryo fibroblasts (DEF) Apoptosis

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Molecular Design of Inhibitors of Efflux pumps in the TolC Outer Membrane Transport Protein

Molecular Design of Inhibitors of Efflux pumps in the TolC O...

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第九届全国计算（机）化学学术会议

作者： Jingyuan Tang~a Ge Zhou~b Hongning Wang~a a bioengineering research center for animal disease prevention and control,School of Life Sciences,sichuan university,chengdu 610064,People’s Republic of china b Faculty of Chemistry,center for Computational Chemistry and Biology,sichuan university,chengdu 610064,People’s Republic of china

The molecule design is carried out as the inhibitor of efflux pumps in the *** indolenine derivatives are selected as the potential inhibitor.A pair of the proton-acceptor and the proton-donor are linked in molecular *** indolenine derivatives are *** Asp and Tyr are linked tightly through the formation of *** by designed molecules at the same *** the tunnel is shrunken and the export of drug is *** the quantum chemistry method,their geometries,atomic charges and interaction energies are studied in detail. The results show that the designed molecules can link Asp and Tyr together by relatively hydrogen *** constrains the tunnel in the distal end of TolC so the drug exporting is *** designed molecules,it is the easiest and the most stable that indo interacts with Asp and Tyr by forming hydrogen *** indo should be the most efficient molecules for shrinking the tunnel and obstruct the export of *** studies showed that these molecules may be a kind of potential and novel inhibitors and our work provide some useful information for further studying them experimentally.

关键词： molecular design inhibitors TolC DFT

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Characterization of codon usage bias in the new discovered DEV dUTPase gene

Characterization of codon usage bias in the new discovered D...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： ZHAO Lichan CHENG Anchun WANG Mingshu CAI Mingsheng YUAN Guiping (1.research center of Avian disease,College of Veterinary Medicine of sichuan Agricultural university,Ya’an 625014,china 2.Key Laboratory of animal disease and Human Health of sichuan Province,Ya’an 625014,china 3.Analytical & Testing center,sichuan university,chengdu,sichuan 610064,china)

A 1344bp gene of Duck Enteritis Virus (DEV) homologous to dUTPase,was first discovered by constructing the DEV genomic *** analyzed the codon usage of DEV dUTPase gene (GenBank accession number DQ486149) by using CAI (Codon Adaptation Index),CHIPS (Codon Heterozygosity in a Protein-coding Sequence) and CUSP (Create a codon usage table) program of EMBOSS (The European Molecular Biology Open Software Suite).The results showed that the DEV dUTPase gene had similar codon usage pattern with avian alpha- and ***-one codons were used in the polypeptide,with a strong bias towards the codons with A and T at the third codon *** of comparison analysis revealed that there are 19 codons showing distinct usage differences between DEV dUTPase and ***,16 between DEV dUTPase and yeast,15 between DEV dUTPase and *** analysis showed high significant differences in DEV dUTPase and yeast (r=0.536,P<0.01).

关键词： DEV dUTPase codon usage bias EMBOSS

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Prediction of secondary structure and B cell epitope for capsid protein of IBV

Prediction of secondary structure and B cell epitope for cap...

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The 15th World Veterinary Poultry Congress

作者： Tai Yang Hongning Wang Sheng Zhou Youfeng Zhang Pengju Zhang Yuling Li Sujun Zhao Lihuan Wang sichuan university “985 project” Science Innovative Platform for Resourceand Environment Protection of Southwestern china. sichuan university bioengineering research center for animal disease prevention and control sichuan province 610064

＜正＞Introduction Infectious bronchitis virus（IBV）continues to cause disease in poultry flocks around the world. Although commercial poultry flocks are routinely vaccinated for IBV,outbreaks of infectious bronchitis still happen due to naturally variant viruses that continue to *** computational methods used in vaccine

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Dynamic Changes of Apoptosis in DEF Cells induced by New Type Gosling Viral Enteritis Virus

Dynamic Changes of Apoptosis in DEF Cells induced by New Typ...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Chen Shun Cheng Anchun Wang Mingshu Chen Xiaoyue (1.Avian disease research center,College of animal Science and Veterinary Medicine of sichuan Agricultural university, 2.Key laboratory of animal disease and Human Health of sichuan Province,Yaan,sichuan,625014,china 3.research center of animal Microecological Engineering,sichuan Agricultural university,Yaan,sichuan,625014,P.R.china)

The monolayer Duck Embryo Fibroblast (DEF) cells were experimentally infected with New Type Gosling Viral Enteritis Virus (NGVEV) and the dynamic changes of apoptosis were detected at different time points after NGVEV infection by using transmission electron microscopy (TEM),DNA agarose gel electrophoresis and Annexin V-FITC/PI stained Fluorescence-Activated Cell Sorter (FACS).The result shows NGVEV can induce infected cells undergo apoptosis and change regularly.A series of characteristic apoptotic morphological changes including shrink of the cells,chromatin condensation and margination,as well as formation of apoptotic bodies were observed by *** typical ladder pattern of DNA fragmentation was demonstrated by agarose gel *** using flow cytometry analysis of Annexin V-FITC/PI staining,the dead,viable,apoptotic and necrotic cells could be analysis quantitatively.

关键词： New Type Gosling Viral Enteritis Virus (NGVEV) Duck Embryo Fibroblasts(DEF) Apoptosis

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Morphological Observation of New Type Gosling Viral Enteritis Virus Virulent Isolate in Infected Duck Embryo Fibroblasts

Morphological Observation of New Type Gosling Viral Enteriti...

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Shun Chen An-Chun Cheng~* Ming-Shu Wang (Key Laboratory of sichuan Provincial animal disease and Human Health,Avian disease research center,College of animal Science and Veterinary Medicine,sichuan Agricultural university 3.research center of animal Microecological Engineering,sichuan Agricultural university,Yaan,sichuan,625014,P.R.china)

The morphogenesis of NGVEV and the characteristic ultrastructural changes of DEF were studied by ultrathin sectioning and transmission electron microscopy after monolayer DEF was infected experimentally with NGVEV virulent *** investigation showed that the typical NGVEV particles were round-shaped with diameter of 75-90 nm and could be found in both the nucleus and the cytoplasm of *** mature virions contained nucleocapsid and nucleic *** virion penetrated into the DEF, replicated and matured in nucleus,and finally was released to the extracellular space via the budding and the disruption of cytoplasmic *** the appearance of progeny NGVEV,some densely electron stained virus-related structures,which were circle-shaped,U-shaped or irregular appearance,could be observed in the cytoplasm of DEF,Three kinds of cytoplasmic inclusion bodies were observed in the infected ***,it was found that NGVEV could induce DEF undergo *** morphological changes of apoptosis included the apoptotic cells shrank,chromatin condensation and margination,as well as the appearance of vacuoles on the cytomembrane aad the formation of apoptotic *** became ultracondensed and aggregated into compact clusters during apoptosis.

关键词： New Type Gosling Viral Enteritis Virus Ultrastructural Changes Morphogenesis Apoptosis Duck Embryo Fibroblasts

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A primarily study on Duck Enteritis Virus (DEV) dUTPase

A primarily study on Duck Enteritis Virus (DEV) dUTPase

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第四届第九次全国学术研讨会暨饲料和动物源食品安全战略论坛

作者： Zhao Li-chan An-chun Cheng Ming-shu Wang Gui-ping Yuan Ren-yong Jia (Key Laboratory of animal disease and Human Health,Avian disease research center,College of Veterinary Medicine,sichuan Agricultural university,Yaan,sichuan,625014,china (Analytical & Testing center,sichuan Univerisity,chengdu,610000,china)

dUTPase is a ubiquitous and important enzyme which hydrolyzes dUTP to *** viruses encode virus specific dUTPase that play an essential role in maintaining the integrity of the viral DNA both by reducing the dUTP levels and by providing the substrate for the thymidylate synthase.A 1344bp gene of Duck Enteritis Virus (DEV),which is homologous to dUTPase,was first identified (GenBank accession number DQ486149) and has been *** gene encodes a protein of 477aa with a predicted molecular mass of 49.7 *** comparisons and multiple alignments showed the conserved motifs of this protein which define the Class 2 dUTPases subfamily with 3-1-2-4-5 motif array,and suggested that DEV dUTPase was quite similar to other identified *** analysis showed that the DEV dUTPase was genetically close to the avian ***,RNA Dot-blot and Western blot,as well as immunofluorescence analysis,indicated that the enzyme is expressed at early and late stages of *** also confirmed that DEV dUTPase localized in the cytoplasm of DEV-infected duck embryo fibroblast as early as 4 hours post infection (hpi),and then the enzyme transferred from cytoplasm to nucleus at 8 hpi then the enzyme expression reached a peak at 12 hpi and it is localized both in the cytoplasm and ***,the expression of DEV dUTPase at both early and late times suggested DEV dUTPase gene might be an early viral gene in DEV vitro infection and played a role in ensuring the fidelity of genome replication.

关键词： Duck Enteritis Virus (DEV) Bioinformatics analysis Transcriptional analysis subcellular localization

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Detection of aminoglycoside-resistance genes in poultry *** isolated from 9 provinces of china by multiplex-PCR

Detection of aminoglycoside-resistance genes in poultry *** ...

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The 15th World Veterinary Poultry Congress

作者： Anyun zhang~1 Hongning wang~2 Wanrong zhou~1 1.College of Resources and Environmental Science sichuan Agricultural university Ya’an 625014. 2.College of Life Science “985 project” science innovative platform for resource and environment protection of southwestern lab of bio-engineering research center for animal disease prevention and control sichuan university chengdu 610064

＜正＞The epidemiology of antimicrobial drug resistance in animals has been widely studied in different coun- tries,and the studies were mainly focused on two important pathogens,Escherichia *** Salmonella spp. The re...

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