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Acta Biochimica et Biophysica Sinica 2006年第3期 213-217页

作者： Guang-Hua HUANG Xin-Yi NIE Jiang-Ye CHEN* State Key laboratory of Molecular biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china

Molecular mechanisms of morphogenesis share many common components between Candidaalbicans and Saccharomyces *** Kss1-associated MAPK cascade and the cAMP/PKA pathwayare two important signal transduction pathways that control morphogenesis in ***.A *** ion-sensing transcription factor gene,CaMAC1,was cloned from *** *** of CaMAC1 in *** promoted filamentous and invasive *** diploid cells,CaMac1could suppress the filamentous growth defect of mutants in the Kss1-associated MAPK pathway and thecAMP/PKA *** haploid strains,ectopic expression of CaMAC1 suppressed the invasive growthdefect of mutants in the MAPK pathway (ste7,ste12 and tec1),but failed to suppress the invasive growthdefect of theflo8 *** results suggest that the activation of CaMac1 is independent of the MAPK andcAMP/PKA pathways in filament formation,but requires Flo8 factor for invasive *** the mediacontaining a high concentration of CuSO,the yeast filamentous and invasive growth was *** effect of CaMac 1 is inhibited by cooper ions.

关键词： CaMac1 MAPK pathway cAMP/PKA pathway morphogenesis Candida albicans Saccharomyces cerevisiae

The in vitro reconstitution of nucleosome and its binding patterns with HMG1/2 and HMG14/17 proteins

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cell Research 2003年第5期13卷 351-359页

作者： SHU BING ZHANG, JIAN HUANG, HUI ZHAO, YI ZHANG, CHUN HuI HOU, XIAO DONG CHENG, CHU JIANG, MIN QIAN LI, JUN HU, RUO LAN QIAN 1 Group of Globin Gene Expression and Regulation, State Key laboratory of Molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china. 2 shanghai institute of Nuclear Research, chinese academy of sciences, shanghai 201800, china Group of Globin Gene Expression and Regulation State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Using atomic force microscopy (AFM), the dynamic process of the in vitro nucleosome reconstitution followed by slow dilution from high salt to low salt was visualized. Data showed that the histone octamers were dissociated from DNA at 1M NaCl. When the salt concentration was slowly reduced to 650 mM and 300 mM, the core histones bound to the naked DNA gradually. Once the salt concentration was reduced to 50 mM the classic 'beads-on-astring' structure was clearly visualized. Furthermore, using the technique of the in vitro reconstitution of nucleosome, the mono- and di- nucleosomes were assembled in vitro with both HS2core (-10681 to -10970 bp) and NCR2 (-372 to -194 bp) DNA sequences in the 5’flanking sequence of human b-globin gene. Data revealed that HMG 1/2 and HMG 14/17 proteins binding to both DNA sequences are changeable following the assembly and disassembly of nucleosomes. We suggest that the changeable binding patterns of HMG 14/17 and HMG1/2 proteins with these regulatory elements may be criti

关键词： atomic force microscopy HMG proteins (HMG1/2 and HMG14/17) human β-globin gene in vitro reconstitution nucleosome.

Cloning and Analyzing of Xenopus Mespo Promoter in Retinoic Acid Regulated Mespo Expression

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Acta Biochimica et Biophysica Sinica 2006年第11期38卷 759-764页

作者： Jin-Hu WANG Xiao-Yan DING* institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,Graduate School of the chinese academy of sciences,shanghai 200031,china instituteofbiochemistryandcellbiology ShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesGraduateSchooloftheChineseAcademyofSciencesShanghai200031China

During vertebrate embryogenesis,presomitic mesoderm cells enter a segmental program togenerate somite,a process termed ***,a member of the bHLH transcription factor family,plays important roles in this ***,how Mespo expression is regulated remains *** this question,we isolated a genomic DNA sequence containing 4317 bp of Mespo 5’ flanking regionin *** assays show that this upstream sequence has transcription *** assayshows that this genomic contig is sufficient to recapitulate the dynamic stage-and tissue-specific expressionpattern of endogenous Mespo from the gastrula to the tailbud *** further mapped a 326 bp DNAsequence responding to retinoic acid *** results shed light on how Mespo expression is regulated,and suggest that retinoic acid signaling pathways play roles in somitogenesis through regulating Mespo.

关键词： bHLH paraxial presomitic segmental tailbud Xenopus laevis

Delay of ZGA initiation occurred in 2-cell blocked mouse embryos

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cell Research 2003年第3期13卷 179-185页

作者： JIA JING QIU,WU WEN ZHANG,ZHI Li WU,YI HONG WANG,MIN QIAN,YI PING LIinstitute,of biochemistry and cell biology,shanghai institutes for biological sciences. chinese academy of sciences,200031 shanghai,chinaSchool of Life Science,East china Normal University. 200062 shanghai,china institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 200031 Shanghai China

One-cell mouse embryos from KM strain and B6C3F1 strain were cultured in M16 medium,in which2-cell block generally *** of KM strain exhibited 2-cell block,whereas B6C3F1 embryos,which are regarded as a nonblocking strain,proceeded to the 4-cell stage in our culture *** is oftenassumed that the block of early development is due to the failure of zygotic gene activation（ZGA）in *** this study we examined protein synthesis patterns by two-dimensional gel electrophoresis of[35S]methionine radiolabeled 2-cell *** from the blocking strain and the nonblocking strainwere compared in their development both in vitro and in *** detection of TRC expression,a markerof ZGA,at 42 h post hCG in KM embryos developed in vitro suggested that ZGA was also initiated even inthe 2-cell arrested ***,a significant delay of ZGA was observed in KM strain as comparedwith normally developed B6C3F1 *** the very beginning of major ZGA as early as 36 h post

关键词： monse embryo two-cell block zygotic gone activation (ZGA).

Expression of Hepatitis C Virus E2 Ectodomain in *** and Its Application in the Detection of Anti-E2 Antibodies in Human Sera

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 57-63页

作者： Jing LIU, Xin-Xin ZHANG, Shen-Ying ZHANG, Min LU, Yu-Ying KONG, Yuan WANG, and Guang-Di LI ( State Key laboratory of Molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, the chinese academy of sciences, shanghai 200031, china 2 Department of Infectious Diseases, Ruijin Hospital, Second Medical University of shanghai, shanghai 200025, china ) State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai 200031 Chin

The second envelope glycoprotein （E2） of hepatitis C virus has been shown to bind human target cells and has become a major target for the development of anti-HCV vaccines. Anti-E2 antibodies have been suggested to be of clinical significance in diagnosis, treatment and prognosis of hepatitis C. However, large-scale expression and purification of E2 proteins in mammalian cells is difficult. As an alternative, E2 fragment （aa 385-730） with a four-amino-acid mutation （aa 568-571 PCNI to RVTS） was expressed as hexa-histidine-tagged full length protein [E2N730（m）] in E. coli and purified to over 85% purity. Purified E2N730（m） was specifically recognized by homologous hepatitis C patient serum in Western blot, suggesting that it displayed E2specific antigenicity. Rabbit antiserum raised against E2N730（m） recognized E2 glyco-proteins expressed in mammalian cells in Western blot. Purified E2N730（m） was used to detect anti-E2 antibodies in human sera and showed better specificity and sensitivity than previousl

关键词： hepatitis C virus envelope protein E2 expression and purification Escherichia coll

Progress in the detection of human genome structural variations

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Science china(Life sciences) 2009年第6期52卷 560-567页

作者： WU XueMei1,3 & XIAO HuaSheng1,2 1 Center of Functional Genomics,Key laboratory of System biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china 2 National Engineering Center for Biochip at shanghai,shanghai 201203,china 3 Graduate School of the chinese academy of sciences,shanghai 200031,china 1. Center of Functional Genomics Key Laboratory of System Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 3. Graduate School of the chinese academy of sciences Shanghai 200031 China 2. National Engineering Center for Biochip at shanghai Shanghai 201203 China

The emerging of high-throughput and high-resolution genomic technologies led to the detection of submicroscopic variants ranging from 1 kb to 3 Mb in the human *** variants include copy number variations(CNVs),inversions,insertions,deletions and other complex rearrangements of DNA *** paper briefly reviews the commonly used technologies to discover both genomic structural variants and their potential ***,we highlight the array-based,PCR-based and sequencing-based assays,including array-based comparative genomic hybridization(aCGH),representational oligonucleotide microarray analysis(ROMA),multiplex amplifiable probe hybridization(MAPH),multiplex ligation-dependent probe amplification(MLPA),paired-end mapping(PEM),and next-generation DNA sequencing ***,we discuss the limitations and challenges of current assays and give advices on how to make the database of genomic variations more reliable.

关键词： structural variation cytogenetic microarray PCR paired end mapping(PEM) next-generation sequencing

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Enhancement of germ cell apoptosis induced by ethanol in transgenic mice overexpressing Fas Ligand

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cell Research 2003年第5期13卷 361-367页

作者： JIA HUA HU, JIE JIANG, YING HUA MA, NA YANG, MAO HU ZHANG, MIN WU, JIAN FEI, LI HE GUO 1 laboratory of Molecular cell biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china 2 College of life science, Zhejiang University, Hangzhou 310027, china 3 shanghai Research Center for Biomodel Organism, shanghai 201203, china 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 2College of life science Zhejiang University Hangzhou 310027 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 2College of life science Zhejiang University Hangzhou 310027 China 3shanghai Research Center for Biomodel Organism Shanghai 201203 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@***

It was suggested that chronic ethanol exposure could result in testicular germ cell apoptosis, but the mechanism is still unclear. In the present study, we use a model of transgenic mice ubiquitously overexpressing human FasL to investigate whether Fas ligand plays a role in ethanol-induced testicular germ cell apoptosis. Both wild-type (WT) mice and transgenic (TG) mice were treated with acute ethanol (20％ v/v) by introperitoneal injection for five times. After ethanol injection, WT mice displayed up-regulation of Fas ligand in the testes, which was shown by FITC-conjugated flow cytometry and western blotting. Moreover, TG mice exhibited significantly more apoptotic germ cells than WT mice did after ethanol injection, which was demonstrated by DNA fragmentation, PI staining flow cytometry and TUNEL staining. In addition, histopathological examination revealed that degenerative changes of epithelial component of the tubules occurred in FasL overexpressing transgenic mice while testicular morphology was

关键词： Fas ligand ethanol apoptosis testes transgenic mouse.

Identification and Characterization of GH Receptor and Serum GH-binding Protein in chinese Sturgeon (Acipenser sinensis)

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 811-816页

作者： Zhi-Yong LIAO1,2 and Shang-Quan ZHU2* 1College of Life sciences, Jiangxi Normal University, Nanchang 330027, china 2Key laboratory of proteomics biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china College of Life sciences Jiangxi Normal UniversityNanchang 330027 China Key laboratory of proteomics biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

chinese sturgeon, a kind of cartilage ganoid, has a history of over one billion years and it is called the living fossil of aquatic biology since it keeps some evolutionary trace. Here, we characterized the growth hormone receptor (GHR) and serum growth hormone binding protein (GHBP) of chinese sturgeon. It was shown that GHR was expressed in various tissues, mainly in hepatic, kidney and intestine tissues. GHR on the hepatic membrane has high and specific affinity for bream GH (brGH) and Scatchard analysis of the binding data showed a single class of high affinity binding site with an association constant Ka of 3.1×109 M–1. A specific band around 94 kD was detected by SDS-PAGE in cross-linking studies of membrane receptors. After incubation of chinese sturgeon serum with 125I-brGH, a 125I-brGH-GHBP complex was identified by Sephadex G-75, indicating that in the serum exists GHBP specially binding to brGH.

关键词： chinese sturgeon growth hormone receptor growth hormone binding protein ligand binding

Gene expression profile favoring phenotypic reversion: a clue for mechanism of tumor suppression by NF-IL6 3'UTR

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cell Research 2003年第6期13卷 509-514页

作者： DING GAN LIU, QIU HONG JIANG, YUN Yl WEI, LI SUN, BEI BEI FU, Fu KUN ZHAO, QIONG ZHOU ’ State Key laboratory of Molecular biology and Key laboratory of proteomics, institute of biochemistryand cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai200031, china. E-mail: dgliu@*** State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. dgliu@***

Transfection of cDNA in 3’untranslated region of human nuclear factor for interleukin-6 (NF-IL6 3’UTR) induced tumor suppression in a human hepatoma cell line. cDNA array analysis was used to reveal changes in gene expression profile leading to tumor suppression The results indicate that this suppression was not due to activation of dsRNA-dependent protein kinase, nor to inactivation of oncogenes; rather, all the changes in expression of known genes, induced by NF-IL6 3’UTR cDNA may be ascribed to the suppression of cellular malignancy. Therefore, our results imply that this 3’untranslated region may have played role of a regulator of gene expression profile.

关键词： cDNA array NF-IL6 hepatoma 3 ’UTR tumor suppression.

GABA transporter 1 transcriptional starting site exhibiting tissue specific difference

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cell Research 2001年第2期11卷 161-163页

作者： JIN XIAO PING, FANG HUANG, NA YANG, BU FENG LU, JIAN FEI, LI HE GUO institute of biochemistry and cell biology, shanghai institutes for biological Science, chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china institute of biochemistry and cell biology Chinese Academy of Sciences Shanghai.

GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address the question, first, 5’ Rapid amplification of cDNA end (RACE) was used to determine GAT1 transcriptional starting sites in neonatal mouse cerebral cortex and intestine, adult mouse brain and adult rat testis. The products of 5’RACE were confirmed by DNA sequencing. We found that the transcript of GAT1 in neonatal mouse cerebral cortex and adult mouse brain starts at the same site (inside of exon 1), while in mouse intestine, GAT1 starts transcription in intron 1, and in rat testis, the transcript of GAT1 has an additional untranslation exon to the 5’ direction.

关键词： GABA transporter 1(GAT1) transcriptional starting site 5’ RACE tissue spedcific difference DNA sequencing.