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Identification and Characterization of glnA Promoter and its Corresponding Trans-regulatory Protein GlnR in the Rifamycin SV Producing Actinomycete,Amycolatopsis mediterranei U32

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Acta Biochimica et Biophysica Sinica 2006年第12期38卷 831-843页

作者： Hao YU~(1&) Wen-Tao PENG~1 Yang LIU~1 Ting WU~2 Yu-Feng YAO~1 Ming-Xue CUI~1 Wei-Hong JIANG~1 Guo-Ping ZHAO~(1,3,4*) 1 Laboratory of Molecular Microbiology,Institute of Plant Physiology and Ecology,shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,shanghai 200032,china 2 Department of Medicinal Chemistry,School of Pharmacy,fudan university,shanghai 200032,china 3 State Key Laboratory of genetic Engineering & Department of Microbiology,School of Life Science,fudan university,shanghai 200433,china 4 Laboratory of Disease and Health Genomics,Chinese National Human Genome center at shanghai,shanghai 201203,china Laboratory of Molecular Microbiology Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China Department of Medicinal Chemistry School of Pharmacy Fudan University Shanghai 200032 China State Key Laboratory of genetic Engineering & Department of Microbiology School of Life Science Fudan University Shanghai 200433 China Laboratory of Disease and Health Genomics Chinese National Human Genome Center at Shanghai Shanghai 201203 China

The genetic requirements for the transcription of glnA,encoding the major glutaminesynthetase in a rifamycin SV-producing Amycolatopsis mediterranei strain,U32,were *** experiments showed that the promoter of U32 glnA （pglnA） was likely to have two transcriptioninitiation sites:P1and P2,located 157 and 45 nucleotides （nt） upstream of the translational start codon,*** mobility shift and DNase I footprinting analyses revealed a 30 bp cis-element locatedat 45 to 75 nt downstream of P1,or 38 to 68 nt upstream of *** sequence of the cis-element displayedhigh similarity to the corresponding regions of pglnA from Streptomyces coelicolor and *** as a reporter gene,the expression levels of U32 pglnA and its deletion derivatives under differentnitrogen-source conditions were analyzed by detecting the catechol dioxygenase activities in ***54,*** J508 and *** FS10 （glnR mutant）.These in vivo studies showed that theactivation of U32 pglnA in *** required GlnR,and its binding to the U32 pglnA was furtherconfirmed by the gel mobility shift *** and heterologous expression of the U32 glnR allowed usto detect the in vitro interaction between the U32 GlnR and the corresponding pglnA *** shown by in vivo glnR inactivation and complementation indicated that GlnR is essential for theactive transcription of glnA in U32.

关键词： glnA promoter cis-element glnR Amycolatopsis mediterranei

Increased Connexin 43 Expression Improves the Migratory and Proliferative Ability of H9c2 Cells by Wnt-3a Overexpression

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Acta Biochimica et Biophysica Sinica 2007年第6期39卷 391-398页

作者： Xiaoyu LIU~(1#), Wen LIU~(1#), Ling YANG~(1#), Beili XIA~1, Jinyan LI~1, Ji ZUO~(1*), and Xiaotian LI~(2*) 1 Department of Celhdar and genetic Medicine, shanghai Medical College. fudan university, shanghai 200032, china 2 The Obstetrics and Gynecology Hospital of the Medical center of fudan university, shanghai 200032. china Department of Cellular and genetic Medicine Shanghai Medical College Fudan UniversityShanghai 200032 China The Obstetrics and Gynecology Hospital of the Medical center of fudan universityshanghai 200032 China

The change of connexin 43 （Cx43） expression and the biological behaviors of Cx43 in rat heart cell line H9c2, expressing Wnt-3a （wingless-type MMTV integration site family, member 3A） were evaluated in the present study. Plasmid pcDNA3.1/Wnt-3a was constructed and transferred into H9c2 cells. The cell model Wnt-3a+-H9c2 steadily expressing Wnt-3a was obtained. Compared with H9c2 and pcDNA3.1-H9c2 cells, the expression of Cx43 in Wnt-3a+-H9c2 cells was clearly increased, the proliferation of Wnt-3a+-H9c2 cells was significantly changed, and cell migration abilities were also improved （P43 expression and altered migratory and proliferative activity in H9c2 cells. Cx43 might be one of the downstream target genes regulated by Wnt-3a.

关键词： Wnt-3a Cx43 H9c2 cell line cell migration

Bacterial Type Ⅰ Glutamine Synthetase of the Rifamycin SV Producing Actinomycete,Amycolatopsis mediterranei U32,is the Only Enzyme Responsible for Glutamine Synthesis under Physiological Conditions

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Acta Biochimica et Biophysica Sinica 2006年第12期 821-830页

作者： Wen-Tao PENG~(1&) Jin WANG~1 Ting WU~2 Jian-Qiang HUANG~1 Jui-Shen CHIAO~1 Guo-Ping ZHAO~(1,3,4) 1 Laboratory of Molecular Microbiology,Institute of Plant Physiology and Ecology,shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,shanghai 200032,china 2 Department of Medicinal Chemistry,School of Pharmacy,fudan university,shanghai 200032,china 3 State Key Laboratory of genetic Engineering/Department of Microbiology,School of Life Science,fudan university,shanghai 200433,china 4 Laboratory of Health and Disease Genomics,Chinese National Haman Genome center at shanghai,shanghai 201203,china

The structural gene for glutamine synthetase,glnA,from Amycolatopsis mediterranei U32 wascloned via screening a genomic library using the analog gene from Streptomyces *** clone wasfunctionally verified by complementing for glutamine requirement of an Escherichia coli glnA null mutantunder the control of a lac *** analysis showed an open reading frame encoding a protein of466 amino acid *** deduced amino acid sequence bears significant homologies to other bacterialtype Ⅰ glutamine synthetases,specifically,71% and 72% identical to the enzymes of *** and Myco-bacterium tuberculosis,*** of this glnA gene in *** U32 led to glutamineauxotrophy with no detectable glutamine synthetase activity in *** contrast,the cloned glnA+gene cancomplement for both phenotypes in *** thus suggested that in *** U32,the glnA geneencoding glutamine synthetase is uniquely responsible for in vivo glutamine synthesis under our laboratorydefined physiological conditions.

关键词： glnA glutamine synthetase Amycolatopsis mediterranei rifamycin SV

Cloning and molecular characterization of a novel lectin gene from Pinellia ternata

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Cell Research 2003年第4期13卷 301-308页

作者： JIAN HONG YAO, XIU YUN ZHAO, ZHI HUA LIAO, JUAN LIN, ZHONG HAI CHEN, FEI CHEN, JUN SONG, XIAO FEN SUN, KE XUAN TANG State Key Laboratory of genetic Engineering, School of Life Sciences, Morgan- Tan International center for Life Sciences, fudan-SJTU-Nottingham Plant Biotechnology R and D center, fudan university, shanghai 200433, chinafudan-SJTU-Nottingham Plant Biotechnology R and D center, Plant Biotechnology Research center, School of Agriculture and Biology, shanghai Jiaotong university, shanghai 200030, china State Key Laboratory of genetic Engineering School of Life Sciences Morgan-Tan International Center for Life Sciences Fudan-SJTU-Nottingham Plant Biotechnology R and D Center Fudan University Shanghai China fudan-SJTU-Nottingham Plant Biotechnology R and D center Plant Biotechnology Research Center School of Agriculture and Biology Shanghai Jiaotong University Shanghai China

The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was succ

关键词： araceae cDNA cloning lectin Pinellia ternata RACE-PCR.

Fluoroquinolone Resistance and Mutation Patterns in gyrA and parC Genes in Neisseria gonorrhoeae Isolates from shanghai, china

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Journal of Huazhong university of Science and Technology(Medical Sciences) 2009年第1期29卷 29-34页

作者：张铁军周晓明 Yue CHEN 顾伟鸣张涛姜庆五 Department of epidemiology School of Public Health Fudan University Key Laboratory of Public Health Safety Ministry of Education Shanghai 200032 China fudan university Affiliated shanghai Public Health center Institute of Preventive Medicine Shanghai 200030 China Department of epidemiology and Community Medicine Faculty of Medicine University of Ottawa Ottawa K5H6 Canada shanghai Skin Disease and STD Hospital Shanghai 200050 China

In order to study the resistance of Neisseria （N.） gonorrhoeae to the fluoroquinolone and detect mutation patterns of quinolone resistance-determining regions （QRDRs） of clinical isolates in shanghai, china, a total of 80 clinical isolates of N. gonorrhoeae were consecutively collected from shanghai. The MIC of fluoroquinolone for the isolates was examined by using the agar dilution method and the mutation profiles of the QRDRs of gyrA and parC were analyzed by sequencing and restriction fragment length polymorphism （RFLP）. Chi-square test was used for comparison of the t：nutation patterns. The results showed that：（1） High percentages of the 8 isolates were resistant to ciprofloxacin （95.0%）, ofloxacin （95.0%） and lomefloxacin （97.5%）, only one strain was susceptible to the ciprofloxacin. （2） Sensitive strains had a substitute of Asp95→Ala in the gyrA, and all isolates that were resistant or intermediated to the ciprofloxacin, had a double mutation in the gyrA （Ser91, Ala 92 and Asp95）. Some strains also had a mutation in the parC. （3） The MICs of these isolates were significantly associated with the mutation patterns in the gyrA and parC. A double mutation of gyrA combined with parC87 mutation was a predominant pattern in shanghai and could mediate high level resistance to ciprofloxacin. It suggests that mutations in the QRDRs of gyrA and parC may be responsible for the fluoroquinolone resistance. And fluoroquinolone could not be used as the first line antibiotics for gonorrhea treatment any more in shanghai, china.

关键词： Neisseria gonorrhoeae resistance fluoroquinolone gyrA parC

维普期刊数据库

Relationships of Aegilops tauschii revealed by DNA fingerprints:The evidence for agriculture exchange between china and the West

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Progress in Natural Science:Materials International 2008年第12期18卷 1525-1531页

作者： Huiting Wei~a Jun Li~a Zhengsong Peng~b Baorong Lu~c Zhijun Zhao~d Wuyun Yang~a a Crop Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,china b Life Science College,china West Normal university,Nanchong 637002,china c The Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering,Institute of Biodiversity Science,fudan university,shanghai 200433,china d Chinese Academy of Social Science,Beijing 110710,china Crop Research Institute Sichuan Academy of Agricultural Sciences Chengdu 610066 China Life Science College China West Normal University Nanchong 637002 China The Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering Institute of Biodiversity Science Fudan University Shanghai 200433 China Chinese Academy of Social Science Beijing 110710 China

genetic diversity and relationships of wild goat grass (Aegilops tauschii Cosson) from Iran and Xinjiang,west china,as well as its weedy type from the Yellow River region of Shaanxi and Henan provinces in china were analyzed by simple sequence repeat (SSR) fingerprints.A high level of genetic diversity in Aegilops tauschii accessions from Iran was observed,and the richness of genetic diversity was followed by accessions from Shaanxi,Henan,and *** weedy type of Aegilops tauschii showed a close genetic relationship with the wild type from different regions in *** results indicated that the weedy Aegilops tauschii found in the Yellow River region was most likely introduced from Iran—the diversity center of Aegilops *** weedy Aegilops tauschii populations found in the Yellow River region may be brought into the central part of china as a weed species together with common wheat (Triticum aestivum L.) from the West during various periods of time in *** finding has provided strong evidence for the introduction of common wheat from the West into china via the Silk Road,and also demonstrated the important role of the Silk Road in the exchange of agri- culture and other relevant technologies between china and the West.

关键词： The Silk Road Aegilops tauschii Common wheat SSR fingerprints genetic diversity Affinity Crop dispersion

Association of DRD2 polymorphisms and chlorpromazine-induced extrapyramidal syndrome in Chinese schizophrenic patients

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Acta Pharmacologica Sinica 2006年第8期27卷 966-970页

作者： Sheng-nan WU~(2,3,5,6) Rui GAO~(2,3,6) Qing-he XING~(2,3,6) Hua-fang LI~4 Yi-feng SHEN~4 Niu-fan GU~4 Guo-yin FENG~4 Lin HE~(2,3,7)2 Bio-X Life Science Research center,shanghai Jiaotong university,shanghai 200030,china 3 Institute of Nutritional Sciences,SIBS,Chinese Academy of Sciences,shanghai 200031,china 4 shanghai Institute of Mental Health,shanghai 200030,china 5 shanghai Institute of Hypertension,Ruijin Hospital,Medical School of shanghai Jiaotong university,shanghai 200025,china Bio-X Life Science Research center Shanghai Jiaotong University Shanghai China Institute of Nutritional Sciences SIBS Chinese Academy of Sciences Shanghai China shanghai Institute of Hypertension Ruijin Hospital Medical School of Shanghai Jiaotong University Shanghai China shanghai Institute of Mental Health Shanghai China

Aim:Extrapyramidal syndrome (EPS) is most commonly affected by typical antip-sychotic drugs that have a high affinity with the D2 ***,manyresearch groups have reported on the positive relationship between the geneticvariations in the DRD2 gene and the therapeutic response in schizophrenia pa-tients as a result of the role of variations in the receptor in modulating *** this study,we evaluate the role DRD2 plays in chlorpromazine-induced EPS in schizophrenic ***:We identified seven SNP(singlenucleotide polymorphism) (-141Cins>del,TaqIB,TaqID,Ser311Cys,rs6275,rs6277and TaqIA) in the DRD2 gene in 146 schizophrenic inpatients (59 with EPS and 87without EPS according to the Simpson-Angus Scale) treated with chlorpromazineafter 8 *** alleles of all loci were determined by PCR (polymerase chainreaction).Results:Polymorphisms TaqlD,Ser311Cys and rs6277 were not poly-morphic in the population recruited in the present *** statistical signifi-cance was found in the allele distribution of-141Cins>del,TaqIB,rs6275 andTaqIA or in the estimated haplotypes (constituted by TaqIB,rs6275 and TaqIA) inlinkage disequilibrium between the two ***:Our results did notlend strong support to the view that the genetic variation of the DRD2 gene playsa major role in the individually variable adverse effect induced by chlorpromazine,at least in Chinese patients with *** results confirmed a previousstudy on the relationship between DRD2 and EPS in Caucasians.

关键词： dopamine receptors basal ganglia disease polymorphism chlorpromazine schizophrenia

SHEsis,a powerful software platform for analyses of linkage disequilibrium,haplotype construction,and genetic association at polymorphism loci

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Cell Research 2005年第2期15卷 97-98页

作者： Yong Yong SHI1,2, Lin HE2,3,*1Bio-X Life Science Research center, shanghai Jiao Tong university, shanghai, china 2Institute for Nutritional Sciences, shanghai Institute of Biological Sciences, Chinese Academy of Sciences, shanghai,china 3NHGG, Bio-X center, shanghai Jiao Tong university, shanghai, china Bio-X Life Science Research center Shanghai Jiao Tong University Shanghai China Institute for Nutritional Sciences Shanghai Institute of Biological Sciences Chinese Academy of Sciences Shanghai China NHGG Bio-X Center Shanghai Jiao Tong University Shanghai China

In multiloci-based genetic association studies of complex diseases, a powerful and high efficient tool for analyses oflinkage disequilibrium (LD) between markers, haplotype distributions and many chi-square/p values with a large numberof samples has been sought for long. In order to achieve the goal of obtaining meaningful results directly from raw data,we developed a robust and user-friendly software platform with a series of tools for analysis in association study withhigh efficiency. The platform has been well evaluated by several sets of real data.

关键词： software linkage disequilibrium haplotype analysis genetic association study.

Comparisons of Graph-structure Clustering Methods for Gene Expression Data

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Acta Biochimica et Biophysica Sinica 2006年第6期38卷 379-384页

作者： Zhuo FANG~1,Lei LIU~(2,4),Jiong YANG~3,Qing-Ming LUO~(1*),and Yi-Xue LI~(2*) ~1 Hubei Bioinformatics and Molecular Imaging Key Laboratory,College of Life Science and Technology, Huazhong university of Science and Technology,Wuhan 430074,china ~2 shanghai center for Bioinformatics Technology,shanghai 200235,china ~3 Department of EECS,Case Western Reserve university,Cleveland 44106,USA ~4 W.M.Keck center for Comparative and Functional Genomics,university of Illinois at Urbana-Champaign,Urbana 61801,USA Hubei Bioinformatics and Molecular Imaging Key Laboratory College of Life Science and Technology Huazhong University of Science and TechnologyWuhan 430074 China shanghai center for Bioinformatics Technologyshanghai 200235 China W.M. Keck center for Comparative and Functional Genomics University of Illinois at Urbana-ChampaignUrbana 61801 USA Department of EECS Case Western Reserve UniversityCleveland 44106 USA

Although many numerical clustering algorithms have been applied to gene expression dataanalysis,the essential step is still biological interpretation by manual *** correlation betweengenetic co-regulation and affiliation to a common biological process is what biologists ***,weintroduce some clustering algorithms that are based on graph structure constituted by biological *** applying a widely used dataset,we compared the result clusters of two of these algorithms in terms ofthe homogeneity of clusters and coherence of annotation and matching *** results show that theclusters of knowledge-guided analysis are the kernel parts of the clusters of Gene Ontology (GO)-Clustersoftware,which contains the genes that are most expression correlative and most consistent with ***,knowledge-guided analysis seems much more applicable than GO-Cluster in a largerdataset.

关键词： clustering expression pattern biological function