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Identification and classification of all potential hemolysin encoding genes and their products from Leptospira interrogans serogroup Icterohaemorrhagiae serovar Lai

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Acta Pharmacologica Sinica 2005年第4期26卷 453-461页

作者： Yi-xuan ZHANG~(2.3,4,5) Yan GENG(2.3,4,5) Bo BI~(2.4) Jian-yong HE~4 Chun-fu WU~4 Xiao-kui GUO~3 Guo-ping ZHAO~(2.4,6,7) 2.Research center of Biotechnology,{2. Institutes for Biological Sciences,Chinese Academy of Sciences,{2. 2.02.3,china.3 Department of Microbiology and Parasitology,{2. Second Medical University,{2. 2.002.,china.4 The School of Pharmaceutical engineering,Shenyang Pharmaceutical University,Shenyang 110016,china.6 Chinese National Human Genome center at {2.,{12. high tech park,{2. 2.12.3,china Research center of Biotechnology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Department of Microbiology and Parasitology Shanghai Second Medical University Shanghai China The School of Pharmaceutical engineering Shenyang Pharmaceutical University Shenyang China Chinese National Human Genome center at shanghai ZhangJiang High Tech Park Shanghai China

Aim:To identify and classify all potential hemolysin candidates of Leptospirainterrogans serogroup Icterohaemorrhagiae serovar ***:All of thepotential hemolysin encoding genes were characterized in *** geneswere cloned and expressed in Escherichia *** hemolytic activities of theexpressed proteins were assayed observing the hemolysis on sheep blood *** activities of the hemolysin candidates were measuredby thin-layer chromatography(TLC)and HPLC for *** and secretion of the hemolysins in L interrogans were studied byreverse transcription polymerase chain reaction,Western blot,and enzyme-linkedimmunosorbent *** and Conclusion:The hemolytic activities ofhemolysin candidates(LA032.,LA0378,LA102.,LA102.,LAI650,LA3050,LA3937,LA4004)from L interrogans strain Lai were *** were furtherdivided into two groups,sphingomyelinase hemolysins and non-sphingomyelinasehemolysins,based on their ability to hydrolyze *** of thesehemolysins were actually expressed in living L interrogans and some of themwere secreted into the *** study establishes an essential findcomplete basis for further studying the contribution of hemolysins to the patho-genesis of L interrogans.

关键词： hemolysin Leptospira interrogans sphingo myelinase

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Comparative genomic hybridization (CGH) microarray reveals genetic diversity among the main epidemic leptospira interrogans in china

Comparative genomic hybridization (CGH) microarray reveals g...

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首届全国微生物基因组学学术研讨会

作者： Ping He, Yue-Ying Sheng, Yao-Zhou Shi, Xiu-Gao Jiang, Jin-Hong Qin, Zhi-Ming Zhang, Guo-Ping Zhao, Xiao-Kui Guo Department of microbiology and parasitology. shanghai Jiaotong university, shanghai 2.02.,{2.National engineering center for biochip at shanghai, zhangjiang high tech park, 2.12.3,{2.

Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Comparative genomic hybridization (CGH) microarray analysis was used to compare the gene content of L. interrogans serovar Lai strain Lai with the gene contents of other L. interrogans strains. Missing genes in a tested strain were detected on a microarray slide spotted with 3,52. open reading frames (ORFs) of L. interrogans serovar Lai strain Lai. For 11 strains subjected to the CGH microarray analyses 2.0 ORFs were found to be absent in at least one strain. The common backbone of the L. interrogans genome was estimated to contain about 2.948 ORFs. The strain-specific genes include genes encoding cell surface structures, metabolism genes and Cellular processes. Genes involved in O-antigen synthesis were highly divergent in strains belonging to the different serogroups, which revealed that antigenic differences between the serogroups are due to genes in the rfb locus. Examination of the genomic context of the strain-specific genes suggests that many are horizontally acquired. The data demonstrated that the CGH microarray technique is useful for genomic comparisons.

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Evaluation of gel entrapped human and rat hepatocytes for the prediction of in vivo metabolic clearance

Evaluation of gel entrapped human and rat hepatocytes for th...

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第九届全国药物和化学异物代谢学术会议

作者： QIU Hong-xia~1,YIN Jian~2.MENG Qin~2.1 Roche R&D center(china. Ltd,72. Cai Lun Road,{2. Hi-tech park,shanghai 2.12.3,china.2.College of Materials Science and Chemical engineering,Zhejiang University,Hangzhou 31002.,Zhejiang,china

AIM:Gel entrapment culture of primary hepatocytes has been demonstrated to retain the expression and function of metabolic *** objective of the current study was to evaluate the utilization of gel entrapped primary human and rat hepatocytes model to predict in vivo hepatic ***:Twenty model compounds ranging from low(cyclosporine A) to high(testosterone) metabolic liability were selected for evaluation of gel entrapped rat hepatocytes, and four among which(warfarin,clozapine,midazolam,and verapamil) for gel entrapped human hepatocytes. The reaction mixture was incubated for 1,2.4,6,8,12.2.,and 2. hr with the density of 2.10 cells/ml culture medium at an initial concentration of 10μ*** temporal profile of the parent compound concentrations were quantitated using an HPLC/MS/MS *** clearance of the test compounds were calculated from the substrate disappearance rate in hepatocytes using the method by Naritomi Y *** the predicted human and rat hepatic clearance was scaled and calculated using the well-stirred ***-specific binding was not accounted for during the calculation. A correlation analysis was conducted between the predicted hepatic clearance and the reported values of in vivo hepatic clearance in human and ***:Metabolic stability of these test compounds spanned a wide range from 0. 06 ml/min/kg for rifampicin to 50.30 ml/min/kg for testosterone in rat *** compounds with intermediate to high metabolic rate,the predicted hepatic clearance were mostly within 2.fold of the observed values from the literature. The in vitro-in vivo correlation was less significant for compounds with low hepatic *** the slowly metabolized compounds,tolbutamide,warfarin,rifampicin and cyclosporine A,the predicted hepatic clearance differed from in vivo data for about 10- *** might be due to indistinctive substrate depletion during the incubation *** predicted human hepatic clearance was 0.06 ml/m

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Genome-Wide Transcriptional Analysis of temperature shift of L. interrogans serovar lai Strain 56601

Genome-Wide Transcriptional Analysis of temperature shift of...

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首届全国微生物基因组学学术研讨会

作者： Jin-Hong QIN, Yue-Ying SHENG, Zhi-Ming ZHANG, Ping HE, Bao-Yu HU, Yang YANG, Shi-Gui LIU, Guo-Ping Zhao, Xiao-Kui GUO School of Life Science /School of Chemical engineering, Sichuan University, Chengdu 610041, PR china.Department of Microbiology and Parasitology, {2. Jiao Tong University School of Medicine, {2. 2.002., PR china.National engineering center for biochip at {2., 151 libing Road, zhangjiang Hi-tech park, Pudong, {2. 2.12.3, china.

Leptospira interrogans is an important mammalian pathogen. Transmission from an environmental source requires adaptations to a range of new environmental conditions in the organs and tissues of the infected host Since many pathogenic ’ bacteria utilize temperature to discern their environment and regulate the synthesis of appropriate proteins, we investigated the effects of temperature on protein synthesis in L. interrogans. We used the L. interrogans serovar lai Strain 56601 genome to construct the microarray. DNA microarray analysis was used to investigate the influence of temperature on global gene expression in strain 56601 grown to mid-exponential phase at 2.癈 and 37癈. A total of 112.genes expression varied significantly in expression at 37癈 relative to 2.癈. The differentially expressed genes included ten kinds of genes which included membrane proteins, hemolysins encoded genes, heat shock proteins, lipoproteins, two-component system, chemotaxis and flagellar proteins, transcriptional regulators, transport and binding protein, metabolic related protein and proteins with no known homologue. Real-time reverse transcription-PCR assays confirmed the microarray data. The data delineate the spectrum of temperature-regulated gene expression in an important human pathogen and provide many unforeseen results of pathogenesis investigation.

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Transcription Character of LA0183-LA02.9 fragment of Leptospira interrogans is similar to a prophage

Transcription Character of LA0183-LA0219 fragment of Leptosp...

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首届全国微生物基因组学学术研讨会

作者： Jin-Hong Qin, Zhi-Ming Zhang, Yang Yang, Bao-Yu Hu, Mathieu Picardeau, Shi-Gui Liu Guo-Ping Zhao, Xiao-Kui Guo School of Life Science /Chemical engineering, Sichuan University, Chengdu 610041, PR china.Department of Microbiology and Parasitology, {2. Jiao Tong University School of Medicine, {2. 2.002.,china.National engineering center for biochip at {2., 151 libing Road, zhangjiang Hi-tech park, Pudong, {2. 2.12.3, china. Laboratoire des Spirochetes, Institut Pasteur, 2. rue du docteur Roux, 7572. Paris Cedex 15, France.

The complete genome sequences of Leptospira interrogans, the agent of leptospirosis, have been obtained. Many sequenced bacterial genomes harbored phage-like elements or cryptic prophages. No phage-like element which has been implicated in pathogenesis, serotype conversion and phage immunity had been studied in L interrogans. BLAST searched using known phage-associated sequences with L interrogans whole genome sequence and phage-like fragment was found around LA0183-LA02.9 which included integrate-like gene, repressor-like gene and other functional phage genes. Operon analysis showed that this phage-like fragment was composed of four transcripts. The function of repressor-like gene (LA0195) was tested using EMSA based on promoter sequence of promoter LA0195 and promoter LA0194 which overlapped in intergenic sequence of LA0194 and LA0195. The results showed that LA0195 protein could bind to the promoter sequence. It is not known whether this fragment still function to its host or only .is a record of the prophage relics. There is currently a lively discussion about the relative contribution of vertical and horizontal elements of evolution in bacteria. Further study of this fragment can shed light on the evolution of both prophages of *** and its hosts.

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Genome-wide Analysis of Smad Targets Reveals the Role of BMP Signaling in Embryonic Stem Cell Fate Determination

Genome-wide Analysis of Smad Targets Reveals the Role of BMP...

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第十一次中国生物物理学术大会暨第九届全国会员代表大会

作者： Teng Fei 1# ,Kai Xia 2. ,Zhang Chen 2.,Hua Chen 1 ,Zhongwei Li 1 ,Jianping Zhang 1 ,Bing Zhou 2.,Huasheng Xiao 3 ,Jing-Dong J.Han 2. ,Ye-Guang Chen 1* 1The State Key Laboratory of Biomembrane and Membrane Biotechnology, Department of Biological Sciences and Biotechnology,Tsinghua University,Beijing 100084,{2.2.AS Key Laboratory of Molecular Developmental Biology and center for Molecular Systems Biology,Institute of Genetics and Developmental Biology,the Chinese Academy of Sciences,Beijing 100101,{2.3National engineering center for biochip at shanghai,zhangjiang Hi-tech park, shanghai,2.12.3,china

＜正＞Embryonic stem（ES）cells are under precise control of both intrinsic self-renewal gene regulatory network and extrinsic growth factor-triggered signaling *** external

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In silico and microarray based genomic approach for identification of potential vaccine candidates against Leptospira interrogans

In silico and microarray based genomic approach for identifi...

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首届全国微生物基因组学学术研讨会

作者： Hong-Liang YANG, Yong-Zhang ZHU, Jin-Hong QIN, Ping HE, Guo-Ping ZHAO and Xiao-Kui GUO Department of Microbiology and Parasitology, shanghai Jiao Tong University School of medicine. 2.002. shanghai, PR{2.Institute of Medical Biology, PeKing Union Medical College & Chinese Academy of Medical Sciences. 650118 Kunming, PR{2.School of Chemical engineering and School of Life Science, Sichuan University, 610041 Chengdu, PR{2.Chinese National Human Genome center at shanghai, zhangjiang high tech park, 2.12.3 shanghai, china

Background Currently available vaccines against leptospirosis are low efficacy, have an unacceptable side-effect profile, do not induce long-term protection, in addition, provide no cross-protection against the different serovars of pathogenic leptospira. There is a need to screening vaccine candidate genes encoding surface-exposed proteins in the genome of Leptospira interrogans. The reverse vaccinology, which has been successful in the identification of vaccines candidates, can be used to identifying the potential vaccine candidates against leptospirosis. Although several papers tried to identify the surface-exposed proteins of leptospire, but vaccine candidate genes encoding the surface-exposed proteins that can not only provide cross-protection against the different serovars of pathogenic leptospires but also have relatively high transcription level are still awaited. Results Bioinformatics, comparative genome analysis as well as transcriptional analysis were used for the identification of novel vaccine candidates in the genome of Leptospira interrogans strain #56601. There were 686 genes (14.5%, 686/472.) were predicted encoding surface-exposed proteins by P-CLASSIFIER, currently the most accurate predicting system for subcellular localization in Gram-negative bacteria analysis combined with signal peptides prediction, a-helix transmembrane topology prediction, integral β-barrel outer membrane protein and putative lipoproteins prediction. The DNA microarray of L. interrogans strain #56601 has been constructed for comparative genome hybridization (CGH) and transcriptome analysis at 37癈 in vitro. 307 differential genes were identified in ten pathogenic serogroups by CGH. The numbers of high transcriptional level (cy3 signal ≥342.and cy5 signal ≥363.5 respectively) are 142. genes. The 686 genes subtract interaction set of genes encoding surface-exposed proteins and 307 differential genes, the number of genes were 62.. After that, the interaction set of 62.genes an

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A Novel Class of Caspase-3 Inhibitors:Tetra- and Pentasubstituted Dihydropyrroles Prepared by Multicomponent Reactions

A Novel Class of Caspase-3 Inhibitors:Tetra- and Pentasubsti...

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2.11年全国药物化学学术会议——药物的源头创新

作者： Qiuhua Zhu~(,),Lixin Gao~§,Huanfeng Jiang~(*,),Shuwen Liu~(*,),Jia Li~(*,§),Zhipeng Chen~,Ronghuan Mai~ and Jinghao Li~ School of Chemistry and Chemical engineering,South china.University of {2.nology,381 Wushan Road, Guangzhou 510640,china.School of Pharmaceutical Sciences,Southern Medical University,Guangzhou,510515,china.Chinese National center for Drug Screening,189 Guoshoujing Road,zhangjiang Hi-{2. park,shanghai, 2.12.3,china

＜正＞Apoptosis is an essential physiological process of tissue development and *** pathogenesis of many diseases,such as cancer and Alzheimer’s disease,is connected with aberrantly regulated *** evidences indicate that caspase-3 is a key executioner caspase and that inhibition of its activity could significantly prevented apoptosis in vitro and in ***,

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