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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 2.0433,china 4 laboratory of Disease and Health Genomics,Chinese National Human Genome Center at shanghai,shanghai 2012.3,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

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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 {12. {12./department of microbiology,School of Life Science,fudan university,shanghai 2.0433,china 4 laboratory of Health and Disease Genomics,Chinese National Haman Genome Center at shanghai,shanghai 2012.3,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

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Genomic research for important pathogenic bacteria in china

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Science china(Life Sciences) 2009年第1期52卷 50-63页

作者： YANG RuiFu1,GUO XiaoKui2,YANG Jian3,JIANG YongQiang1,PANG Bo4,CHEN Chen5,YAO YuFeng2,QIN JinHong2 & LI QingTian2 1 laboratory of Analytical microbiology,state key laboratory of Pathogen and Biosecurity,Beijing Institute of microbiology and Epidemiology,Beijing 100071,china 2 department of Medical microbiology and Parasitology,Institutes of Medical Sciences,shanghai Jiao Tong university School of Medicine,shanghai 200025,china 3 state key laboratory for Molecular Virology and genetic engineering,Institute of Pathogen Biology,Chinese Academy of Medical Sciences,Beijing 100176,china 4 state key laboratory for Infectious Disease Prevention and Control,Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,china 5 Beijing Genomics Institute at Shenzhen,Shenzhen 518083,china 1. laboratory of Analytical microbiology State Key Laboratory of Pathogen and Biosecurity Beijing Institute of Microbiology and Epidemiology Beijing 100071 China 2. department of Medical microbiology and Parasitology Institutes of Medical Sciences Shanghai Jiao Tong University School of Medicine Shanghai 200025 China 3. state key laboratory for Molecular Virology and genetic engineering Institute of Pathogen Biology Chinese Academy of Medical Sciences Beijing 100176 China 4. state key laboratory for Infectious Disease Prevention and Control Institute for Communicable Disease Control and Prevention Chinese Center for Disease Control and Prevention Beijing 102206 China 5. Beijing Genomics Institute at Shenzhen Shenzhen 518083 China

Rapid accumulation of bacterial genomic data offered an unprecedented opportunity to understand bacterial biology from a holistic view of *** can thus closely look at the way in which a pathogen is evolved,and these data has been applied to molecular epidemiology and microbial forensics,and screening of novel diagnostic,vaccine and drug *** newly developed high-throughput low-cost sequencing technologies,such as 454,Solexa and SOLiD,will promote the acquisition and application of genomic data in new research areas that we dared not imagine previously,such as the metagenomics of human gastric-intestinal tract,for better and comprehensive understanding of human health and disease.

关键词： genome pathogenic bacteria sequencing application

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cDNA Array Analysis of the Differential Expression Change in Virulence-related Genes During the Development of Resistance in Candida albicans

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Acta Biochimica et Biophysica Sinica 2005年第7期37卷 463-472页

作者： Zheng XU Yong-Bing CAO~1 Jun-Dong ZHANG~1 Ying-Ying CAO~1 Ping-Hui GAO~1 De-Jun WANG~1 Xu-Ping FU~2 Kang YING~2 Wan-Sheng CHEN~1 Yuan-Ying JIANG~1 1 department of Pharmacology,School of Pharmacy,Second Military Medical university,shanghai 2.0433,china 2 state key laboratory of genetic engineering,Institute of genetics.School of Life Science,fudan university,shanghai 2.0433,china departmentofPharmacology SchoolofPharmacySecondMilitaryMedicalUniversityShanghai200433China

Candida albicans is the most frequently isolated fungus in immunocompromised patientsassociated with mucosal and deep-tissue *** investigate the correlation between virulence andresistance on a gene expression profile in ***,we examined the changes in virulence-related genesduring the development of resistance in *** from bone marrow transplant patients using a constructedcDNA array representing 3096 *** addition to the genes known to be associated with azole resistance,16 virulence-related genes were identified,whose differential expressions were newly found to be associatedwith the resistant *** expressions for these genes were confirmed by ***,the up-regulation of EFG1,CPH2,TEC1,CDC24,SAP10,ALS9,SNF1,SPO72and BDF1,and the down-regulation of RAD32,IPF3636 and UBI4 resulted in stronger virulence andinvasiveness in the resistant isolates compared with susceptible *** findings provide a link betweenthe expression of virulence genes and development of resistance during *** infection in bone marrowtransplant (BMT) patients,where *** induces hyphal formation and expression change in multiplevirulence factors.

关键词： Candida albicans cDNA array resistance virulence

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Prokaryotic Expression and Preparation of Polyantibody of Human Histydyl-tRNA Synthetase Related Gene

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Journal of Huazhong university of Science and Technology(Medical Sciences) 2004年第6期24卷 535-536,555页

作者：孟宪芳施静刘晓春陈金中 department of Neurobiology Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 China state key Library of genetic engineering School of Life Science Fudan University Shanghai 200030 China

Summary: The aim of this study was to express and purify human histydyl-tRNA synthetase related gene and to prepare its polyantibody. The open reading frame was amplified by PCR, and then recombined into prokaryotic expression vector pQE30 and transformed into E. coli M15 for expression. The expressed products were induced by IPTG after the reconstructed pQE30 was transferred into M15. After purified by Ni affinity chromatography, the product was identified to be a single band by SDS-PAGE. The rabbits were inoculated with purified products. High-titer polyantibody was successfully prepared. Highly-purified expression product and prepared polyantibody may provide a good basis for further study.

关键词： human histydyl-tRNA synthetase related gene polyantibody purification

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Molecular genetic,cell biological and genomic analyses of meiotic recombination in Arabidopsis

Molecular genetic,cell biological and genomic analyses of me...

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从植物科学到农业发展——2012.国植物生物学大会

作者： Hong Ma state key laboratory of genetic engineering,Institute of Plant Biology,Center for Evolutionary Biology,School of Life Sciences,fudan university,shanghai 2000433,china department of Biology, the Pennsylvania state university,university Park,PA 16802 USA.

Meiotic recombination is crucial for normal meiotic homolog association and segregation, thereby ensuring normal fertility in plants and other *** addition,meiotic recombination redistributes genome variations and affects allelic frequencies each *** have identified and characterized several Arabidopsis genes important for meiotic recombination using molecular *** identify additional genes potentially important for meiotic recombination and other meiotic functions,we analyzed Arabidopsis male meiocyte transcriptome using high throughput sequencing of cDNAs and detected the expression of～16,000 *** found that,among 3,378 known Arabidopsis gene families,3,265 contained meiocyte-expressed genes and 18 were overrepresented in male *** addition,a large number of genes expressed in meiocytes are conserved in other organisms,including 503 single-copy genes shared between Arabidopsis,mouse and *** the genes detected in Arabidopsis meiocytes are those similar to genes for recombination or DNA synthesis,suggesting that they might be involved in meiotic recombination;genetic and cell biological studies of some of these genes reveal new mechanisms of meiotic recombination and support a key role for lagging strand DNA synthesis in crossover formation via the interference-sensitive *** highly conserved nature of these genes further suggests that the newly uncovered mechanisms might be widespread in eukaryotes.

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Analysis of the Transcriptional Networks for Arabidopsis Pollen Development

Analysis of the Transcriptional Networks for Arabidopsis Pol...

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中国植物生理学会第十次会员代表大会暨全国学术年会

作者： Hong Mastate key laboratory of genetic engineering and Institute of Plant Biology, Center for Evolutionary Biology, School of Life Sciences, fudan university, shanghai, china 2.0433 Institutes of Biomedical Sciences, fudan university, shanghai, china 200032 {12. of Biology and the Huck Institutes of the Life Sciences, the Pennsylvania state university, university Park, PA 16802

＜正＞Pollen development occurs in the male organ anther, which has both sporogenous （the germ-line） and somatic

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Receptor-like Kinase and Other Signaling Molecules in Controlling Early Anther Cell Division and Differentiation

Receptor-like Kinase and Other Signaling Molecules in Contro...

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2009整合植物生物学研讨会

作者： Hong Ma~* state key laboratory of genetic engineering and Institute of Plant Biology,Center for Evolutionary Biology,School of Life Sciences,fudan university,shanghai 2.0433,china Institutes of Biomedical Sciences,fudan university,shanghai 200032,china department of Biology and the Huck Institutes of the Life Sciences,the Pennsylvania state {12.,{12. Park,PA16802 USA.

Anther development is crucial for plant reproductive success and its understanding can contribute to better plant breeding *** early anther development,several somatic cell layers and meiocytes are formed by successive rounds of cell division and associated cell *** is clear that various cell types need to be coordinated,but

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NRP proteins exert their histone chaperone role in the form of dimer during transcriptional modulation

NRP proteins exert their histone chaperone role in the form ...

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上海市植物生理学会成立三十周年庆祝活动暨第三届上海市植物生理学青年学术研讨会

作者： Yan Zhu, Juan Gao, Chi Zhang, Aiwu Dong * state key laboratory of genetic engineering, Institute of Plant Biology, department of Biochemistry, School of Life Sciences, fudan university, shanghai 2.0433, PR china

Local chromatin compaction and dynamic modulation are tightly connected with the delicate regulation on specific gene transcription. Considering the crucial roles in the reversible dynamics

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A novel ASYMMETRIC LEAVES1 and 2 function in Arabidopsis floral organ development

A novel ASYMMETRIC LEAVES1 and 2 function in Arabidopsis flo...

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2006年拟南芥研究学术研讨会

作者： Ben Xu Aiwu Dong Hai Huang National laboratory of Plant Molecular genetics, shanghai Institute of Plant Physiology and Ecology, shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, shanghai 200032, china state key laboratory of genetic engineering, department of Biochemistry, School of Life Sciences, {12., {12., china

＜正＞The Arabidopsis ASYMMETRIC LEAVES1 and 2 （AS1 and AS2） genes are demonstrated to repress class I KNOTTED1-like homeobox genes and to promote the leaf adaxial specifying genes PHABULOSA, PHAVOLUTA and REVOLUTA during leaf morphogenesis. However, very little is known about AS1/2 roles in flower development, although as1 and as2 flowers appear not completely normal. To investigate the AS1/2 function in flower development, we constructed double mutants by combining as1 or as2 with

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