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Expression of Overlapping PreS1 Fragment Recombinant Proteins for the Determination of Immunogenic Domains in HBsAg PreS1 Region

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Acta Biochimica et Biophysica Sinica 2004年第6期36卷 397-404页

作者： Wei-Guo HU, Jun WEI&, Xin-Xiu YANG, Heng-Chuan XIA, Feng LI, and Zu-Chuan ZHANG* key laboratory of Proteomics, Institute of Biochemistry and Cell biology, shanghai institutes for biological sciences,the Chinese {4. of sciences, shanghai 2.0031, china key laboratory of Proteomics Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of SciencesShanghai 200031 China

In this study, eight preS1 fragments overlapped in preS1 (21–119) region of HBV adr subtype, i.e. preS1 (21–4.), preS1 (34.59), preS1 (4.–70), preS1 (60–85), preS1 (71–94., preS1 (86–109), preS1 (95–119) and preS1 (21–119), were cloned by PCR, and expressed as GST fusion proteins. These GST- preS1 fusion proteins were highly expressed in soluble form in E. coli, and about 50 to 90 mg soluble fusion proteins were purified from 1 L culture. Using these fusion proteins, the immunogenic domains in preS1 (21– 119) region were identified by Western blot analysis and competitive ELISA. The results showed that the immunogenic domains mainly existed in preS1 (21–59) in N-terminus and preS1 (95–109) in C-terminus, and more importantly, a major immunogenic domain preS1 (34.59), which has much stronger immunogenicity, was identified. It was also supported by the predictions of secondary structure and immunological property in the preS1 (21–119) region. The results here would be helpful for the design of new vaccines against HBV.

关键词： HBV preS1 overlapping fragment immunogenic domain

Overexpression of a novel gene , Cms1, can rescue the growth arrest of a Saccharomyces cerevisiae mcm10 suppressor

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Cell Research 2001年第4期11卷 285-291页

作者： WANG JI WU, JIA RUI WU State key laboratory of Molecular biology, Institute of Biochemistry and Cell biology, {4. Research Center of Life Science, shanghai institutes for biological sciences, Chinese academy of sciences, 320 Yue- Yang Road, {4. 1State key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Research Center of Life Science Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

MCM10 protein is an essential replication factor involved in the initiation of DNA replication. A mcm10 mutan (mcm10-1) of budding yeast shows a growth arrest at 37 C. In the present work, we have isolated a mcm10-1 suppressor strain, which grows at 37 C. Interestingly, this mcm10-1 suppressor undergoes cell Cycle arrest at 14.C. A novel gene, YLR003c, is identified by high-copy complementation of this suppressor. We called it as Cms1 (Complementation of Mcm 10 Suppressor). Furthermore, the experiments of trans- formation show that cells of mcm10-1 suppressor with high-copy plasmid but not low-copy plasmid grow at 14.C, indicating that overexpression of Cms1 can rescue the growth arrest of this mcm10 suppressor at non-permissive temperature. These results suggest that CMS1 protein may functionally interact with MCM10 protein and play a role in the regulation of DNA replication and cell cycle control.

关键词： Saccharomyces cerevisiae DNA replication Mcm10 Cms1 cell cycle.

Effects of Huangqi (Hex) on Inducing Cell Differentiation and Cell Death in K562 and HEL Cells

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 211-217页

作者： Xiao-Dong CHENG1, Chun-Hui HOU1, Xue-Jun ZHANG2, Heng-Yue XIE1, Wei-Ying ZHOU1, Lei YANG2, Shu-Bing ZHANG1, and Ruo-Lan QIAN1 State key laboratory of Molecular biology, Institute of Biochemistry and Cell biology, {4., the Chinese academy of sciences, 320 Yueyang Road, shanghai 2.0031, china laboratory of Molecular Cell biology, Institute of Biochemistry and Cell biology, {4., the Chinese academy of sciences,320 Yueyang Road shanghai 2.0031,china the Chinese academy of sciences, 320 Yueyang Road, shanghai 2.0031, china State key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China laboratory of Molecular Cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China

Huangqi (Astragalus membranaceus), a traditional Chinese medicine, has been used to ameliorate side effects of cancer chemotherapy in china. However, little is known about its molecular mechanisms. Here we show that induction of K562 or HEL cells with 1.5 mg/ml of Huangqi (Hex) (Components extracted from Huangqi) for 3–5 d results in the expression of β-globin gene in both cell lines and leads to terminal differentiation. Moreover, the apoptosis in HEL cells can be induced by increasing concentration of Huangqi (Hex) to 4.5 mg/ml for 3–5 d. Upregulation of Apaf-1, caspase-3 and acetylcholinesterase (AChE) in HEL cells may play a crucial role in the process of apoptosis. The prospect of inducing expression of adult (β) globin gene and apoptosis selectively in cancer cells is obviously attractive from a therapeutic point of view.

关键词： cell death and differentiation Huangqi (Hex) human erythroleukemia cell lines

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 {4. of sciences,shanghai 2.0031,china 2 National Engineering Center for Biochip at shanghai,shanghai 201203,china 3 Graduate School of the Chinese {4. of sciences,shanghai 2.0031,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

A meta-analysis of oxidative stress markers in schizophrenia

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中国科学：生命科学英文版 2010年第1期53卷 112-124页

作者： ZHANG Ming1,2,ZHAO ZhongMing3,HE Lin1,2,4.& WAN ChunLing1,2 1 Bio-X Center,key laboratory for the Genetics of Developmental and Neuropsychiatric Disorders(Ministry of Education),shanghai Jiao Tong University,shanghai 200030,{4. 2 institutes for Nutritional sciences,shanghai Institute of biological sciences,Chinese academy of sciences,shanghai 2.0031,{4. 3 Departments of Biomedical Informatics,Psychiatry,and Cancer {14.,Vanderbilt University Medical Center,Nashville,TN 37232,USA 4.institutes of Biomedical sciences,Fudan University,shanghai 200032,{4. 1. Bio-X Center Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education) Shanghai Jiao Tong University Shanghai 200030 China2. Institutes for Nutritional Sciences Shanghai Institute of Biological Sciences Chinese Academy of Sciences Shanghai 200031 China3. Departments of Biomedical Informatics Psychiatry and Cancer Biology Vanderbilt University Medical Center Nashville TN 37232 USA4. Institutes of Biomedical Sciences Fudan University Shanghai 200032 China

Oxidative stress has been identified as a possible element in the neuropathological processes of schizophrenia(SCZ).Alteration of oxidative stress markers has been reported in SCZ studies,but with inconsistent *** evaluate the risk of oxidative stress to schizophrenia,a meta-analysis was conducted,including five markers of oxidative stress [thiobarbituric reactive substances(TBARS),nitric oxide(NO),catalase(CAT),glutathione peroxidase(GP) and superoxide dismutase(SOD)] in SCZ patients versus healthy *** study showed that TBARS and NO significantly increased in SCZ,while SOD activity significantly decreased in the disorganized type of SCZ *** significant effect size was found for the activities of GP and CAT in SCZ patients(P>0.05).Egger’s regression test observed no significant publication bias across the oxidative stress markers,but found high heterogeneities in all the 5 *** subgroup analysis suggested that the ethnicity,sample size of patients and sample sources may contribute to the heterogeneity of the results for TBARS,NO and *** result further demonstrated the involvement of oxidative stress in the pathophysiology of schizophrenia.

关键词： schizophrenia oxidative stress meta-analysis lipid peroxidation nitric oxide antioxidant enzymes

The contribution of cis-regulatory elements to head-to-head gene pairs' co-expression pattern

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Science china(Life sciences) 2009年第1期52卷 74-79页

作者： HAO Pei1,2,YU Yao2,3,ZHANG XiaoYan4.TU Kang2,FAN HaiWei2 & ZHONG Yang1 1 School of Life sciences,Fudan University,shanghai 2004.3,china 2 Bioinformatics Center,key Lab of {4. biology,shanghai institutes for biological sciences,Chinese academy of sciences,shanghai 2.0031,china 3 shanghai Center for Bioinformation Technology,shanghai 200235,china 4.School of Life sciences,Tongji University,shanghai 200092,china 1. School of Life sciences Fudan University Shanghai 200433 China 2. Bioinformatics Center Key Lab of Systems Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 3. shanghai Center for Bioinformation Technology Shanghai 200235 China 4. School of Life sciences Tongji University Shanghai 200092 China

Transcription regulation is one of the most critical pipelines in biological process,in which cis-elements play the role as gene expression *** attempt to deduce the principles underlying the co-expression of "head-to-head" gene pairs by analyzing activities or behaviors of the shared cis-elements.A network component analysis was performed to estimate the impact of cis-elements on gene promoters and their activities under different *** discoveries reveal how biological system uses those regulatory elements to control the expression pattern of "head-to-head" gene pairs and the whole transcription regulation system.

关键词： transcription regulation cis-elements "head-to-head" gene pairs

miRNAs modulate the drug response of tumor cells

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Science china(Life sciences) 2009年第9期52卷 797-801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 {4. of sciences,shanghai 2.0031,china 2 National Engineering Center for Biochip at shanghai,shanghai 201203,china 3 Graduate University of Chinese {4. of sciences,shanghai 2.0031,china Center of Functional Genomics Key Laboratory of System Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China National Engineering Center for Biochip at shanghai Shanghai 201203 China Graduate University of Chinese academy of sciences Shanghai 200031 China

Chemotherapy is one of the major treatments of malignant carcinomas. However,its efficiency is affected by both intrinsic and acquired resistance to anticancer drugs. The cellular mechanisms of drug resistance include the overexpression of energy-dependent transporters that eject anticancer drugs from cells such as p-glycoprotein and multidrug resistance related protein (MRP),the mutation of drug targets,the activation of DNA repair pathways,the defects in cellular death pathways and so on. The genetic and epigenetic changes of these genes can lead to cancer drug resistance. Among these mechanisms,microRNAs (miRNAs) which are critical and essential for many important processes such as development,differentiation,and even carcinogenesis have been reported to regulate the chemosen-sitivity of tumor cells. In this paper we briefly review the relationship between miRNA and cancer drug resistance.

关键词： cancer drug resistance miRNAs

Identification of glutamate transporters and receptors in mouse testis

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Acta Pharmacologica Sinica 2004年第03期25卷 112-117页

作者： Jia-hua HU,Na YANG2,Ying-hua MA,Jie JIANG,Jin-fu ZHANG3,Jian FEI4.Li-he GUO4.laboratory of Molecular Cell biology,Institute of Biochemistry and Cell biology,shanghai institutes for biological sciences,Chinese {4. of sciences,shanghai 2.0031 3Department of Urology,Tongji Hospital,Tongji University,shanghai 200065,china laboratory of Molecular Cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological Sciences Chinese Academy of Sciences Department of Urology Tongji Hospital Tongji University

AIM: To investigate the presence of glutamate transporters and receptors in mouse testis. METHODS: Glutamate uptake analysis was performed to study the function of glutamate transporters in mouse testis. Comparative RT-PCR technique and sequencing analysis were used to study the expression of glutamate receptors and transport- ers in mouse testis. RESULTS: Mouse testis possessed glutamate uptake capacity with sodium-dependence. Vmax value of glutamate uptake was (1.60 ± 0.21) pmol/min per mg protein and Km value of glutamate uptake was (11.0±1.6) μmol/L in mouse testis according to saturation analysis. Furthermore, the uptake activity could be inhibited by DHK (GLT1 selective inhibitor) and THA (glutamate uptake inhibitor). In addition, RT-PCR results revealed that glutamate transporters (GLT1 and EAAC1) and ionotropic glutamate receptors (NR1, NR2B, GluR6 and KA2) were expressed in mouse testis. CONCLUSION: Glutamate transporters and receptors do exist in mouse testis.

关键词： glutamates receptors amino acid transport systems testis uptake

Six Specific Lysihe Residues are Crucial in Maintaining the Structure and Function of Soluble Manganese Stabilizing Protein

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Acta Biochimica et Biophysica Sinica 2006年第9期 611-619页

作者： Jin-Peng GAO~(1,2#) Feng ZHANG~(1,3#) Li ZHANG~4.Yin-Long GUO~4.Kang-Cheng RUAN~5 De-An JIANG~3* Chun-He XU~1 1 Institute of Plant Physiology and Ecology,shanghai institutes for biological sciences,Chinese academy of sciences,shanghai 200032,{4. 2 Graduate School of the Chinese academy of sciences,shanghai 200032,{4. 3 College of Life sciences,Zhejiang University,Hangzhou 310029,{4. 4.shanghai Mass Spectrometry Center,shanghai Institute of Organic Chemistry,Chinese academy of sciences,shanghai 2.0031,{4. 5 laboratory of Proteomics,Institute of Biochemistry and Cell biology,shanghai institutes for biological sciences,Chinese academy of sciences,shanghai 2.0031,{4.

When manganese stabilizing protein （MSP） was treated with 0.5mM N-succinimidyl propionate（NSP）,the rebinding ability and oxygen-releasing capabilities of the modified MSP were not altered,in spiteof changes of MSP surface Lys ***,far-ultraviolet circular dichroism and intrinsicfluorescence spectra analysis revealed that 0.5 mM NSP-modified MSP retained most of its native secondaryand tertiary *** of the sites of NSP modification by Staphylococcus V8protease digestion ofthe modified protein,as well as analysis by matrix-assisted laser desorption ionization-time of flight massspectrometry,indicated that seven Lys residues were *** results suggested that these residues arenot absolutely essential to the structure and function of ***,when the NSP concentration wasincreased to 4.M,the modified MSP was unable to bind photosystem Ⅱ and completely lost its *** far-ultraviolet circular dichroism and intrinsic fluorescence spectra analysis revealed a clearconformational change in MSP after 4.mM NSP treatment,suggesting that some Lys residues are involved inmaintaining the structure and function of *** by matrix-assisted laser desorption ionization-timeof flight mass spectrometry indicated that another six Lys residues,namely Lys20,Lys101,Lys196,Lys207,Lys130 （or Lys137） and Lys66 （or Lys76）,were modified by 4.mM ***,these six Lys residuesare crucial in maintaining the structure and function of soluble MSP.

关键词： chemical modification fluorescence spectrum circular dichroism spectrum manganese stabilizing protein Lys residue