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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 234-240页

作者： Jin-Guo DING~(1,3) Jian FEI~2 Da-Fu CUI~1 You-Shang ZHANG~1 ~1Institute of Biochemistry and Cell Biology,and ~2Model Organism Research Center,shanghai Institutes for Biologicol sciences, chinese academy of sciences,shanghai 200031,china ~3graduate school of the chinese academy of sciences,shanghai 200031,china Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China graduate school of the chinese academy of sciences shanghai 200031 China Model Organism Research Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Monomeric B27 Lys destripeptide insulin (B27 Lys DTrI) was designed and produced from itsprecursor expressed in Pichia pastoris through tryptic hydrolysis instead of the less efficient *** monomeric B27 Lys DTrI precursor (MIP) was purified from a cultured medium *** by a combination of hydrophobic,size-exclusion,and ion-exchange *** purifiedMIP was converted,by tryptic hydrolysis,to B27 Lys DTrI,which was then purified by ion-exchangechromatography to homogeneity as assessed by native gel electrophoresis,HPLC,amino acid composition,and electrospray mass-spectrometric analysis.B27 Lys DTrI exhibited superior monomeric properties insize-exclusion *** yield of MIP was 200 mg per liter of culture,and the overall yield ofpurified B27 Lys DTrI from the crude MIP was 70%.The in vivo biological activity of B27 Lys DTrI asdetermined by the mouse convulsion assay was 21 U/mg,identical to that obtained by semisynthesis.

关键词： monomeric B27 Lys destripeptide insulin tryptic hydrolysis Pichia pastoris

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 Center of Functional Genomics Key Laboratory of System Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China graduate school of the chinese academy of sciences Shanghai China National Engineering Center for Biochip at shanghai Shanghai 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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Receptor tyrosine kinases positively regulate BACE activity and Amyloid-β production through enhancing BACE internalization

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Cell Research 2007年第5期17卷 389-401页

作者： Lin Zou1,2 Zhu Wang1 Li Shen1,3 Guo Bin Bao1 Tian Wang1 Jiu Hong Kang1 Gang Pei1 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 Clinical Molecular Medical Center, Children's Hospital, Chongqing University of Medical Sicences, Chongqing 400011, china 3 graduate school of the chinese academy of sciences, shanghai 200031, china Laboratory of Molecular Cell Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai China Clinical Molecular Medical Center Children's Hospital Chongqing University of Medical Sicences Chongqing China graduate school of the chinese academy of sciences Shanghai China

Amyloid-β (AP) peptide, the primary constituent of senile plaques in Alzheimer's disease (AD), is generated byβ-secretase- and γ-secretase-mediated sequential proteolysis of the amyloid precursor protein (APP). The aspartic pro-tease, β -site APP cleavage enzyme (BACE), has been identified as the main P-secretase in brain but the regulation of itsactivity is largely unclear. Here, we demonstrate that both BACE activity and subsequent Aβ production are enhancedafter stimulation of receptor tyrosine kinases (RTKs), such as the receptors for epidermal growth factor (EGF) and nervegrowth factor (NGF), in cultured cells as well as in mouse hippocampus. Furthermore, stimulation of RTKs also inducesBACE internalization into endosomes and Golgi apparatus. This enhancement of BACE activity and Aβ production uponRTK activation could be specifically inhibited by Src family kinase inhibitors and by depletion of endogenous c-Src withRNAi, and could be mimicked by over-expressed c-Src. Moreover, blockage of BACE internalization by a dominantnegative form of Rab5 also abolished the enhancement of BACE activity and Aβ production, indicating the requirementof BACE internalization for the enhanced activity. Taken together, our study presents evidence that BACE activity andAβ production are under the regulation of RTKs and this is achieved via RTK-stimulated BACE internalization, andsuggests that an aberration of such regulation might contribute to pathogenic Aβ production.

关键词： β-site APP cleavage enzyme RTK Amyloid-β Src

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 Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

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

The cDNA and Genomic DNA Organization of a Novel Toxin SHT-I from Spider Ornithoctonus huwena

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 656-660页

作者： Peng QIAO, Xiao-Pan ZUO, Zhi-Fang CHAI, and Yong-Hua JI* Institute of Physiology, shanghai Institutes for Biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china Institute of Physiology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

In present study the full-length cDNA of a novel toxin from the venom gland of spider Ornithoctonus huwena, named as SHT-I, has been cloned using the strategy of rapid amplification of cDNA ends, and then the whole genomic sequence of SHT-I (Selenocosmia huwena toxin-I) was determined using sequence-specific primers synthesized based on the acquired 3’ and 5’ ends of SHT-I cDNA sequence. It is unexpectedly found that intron was lacking in the genomic sequence of SHT-I. The result might evoke an interesting question whether the gene code of inhibitor cystine-knot peptides from spider venom is distinct from that of those known toxic peptides of scorpion and cone snail.

关键词： Ornithoctonus huwena spider cDNA genomic DNA peptide toxin ICK motif

Association between Alcohol, Smoking and HLA-DQA1*0201 Genotype in Psoriasis

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 597-602页

作者： Guang-Yong ZHENG1,2#, Sheng-Cai WEI3#, Tie-Liu SHI1, and Yi-Xue LI1* 1Bioinformatics Center, shanghai Institutes for Biological sciences, chinese academy of sciences, graduate school of the chinese academy of sciences, shanghai 200031, china Research Center for Proteome Analysis, 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 3Guangzhou Institute of Dermatology, Guangzhou 510095, china Bioinformatics Center Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Research Center for Proteome Analysis Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China Guangzhou Institute of DermatologyGuangzhou 510095 China

Psoriasis is a chronic skin disease triggered by genetic, environment or other risk factors such as infection, drugs, stress, moisture, alcohol, and smoking. A major psoriasis susceptibility locus at 6p21.3 has been identified. Further studies found that HLA-DQA1*0201 allele was associated with psoriasis. However, there were few data exploring an association between the environmental factors and susceptibility genes. In this study, the samples of 189 patients with psoriasis and 333 healthy controls were collected with their consent and were carried on analysis through polymerase chain reaction sequence-specific primer (PCR- SSP) method. The proportion of male psoriasis patients engaging in the smoking and alcohol was much higher than that of the control group (P6.91, P<1.0×10–4).

关键词： psoriasis HLA-DQA1*0201 allele smoking alcohol association

Hyaluronan Promotes Tumor Lymphangiogenesis and Intralymphantic Tumor Growth in Xenografts

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Acta Biochimica et Biophysica Sinica 2005年第9期37卷 601-606页

作者： Li-Xia GUO Ke ZOU Ji-Hang JW Hong XIE Laboratory of Biotherapy,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 Laboratory of Biotherapy Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

Hyaluronan (HA),a high molecular weight glycosaminoglycan in the extracellular matrix,hasbeen implicated in the promotion of malignant phenotypes,including tumor ***,little isknown about the effect of HA on tumor-associated *** this study,mouse hepatocellularcarcinoma Hca-F cells combined with or without HA were injected subcutaneously into C3H/Hej mice,thenangiogenesis and lymphangiogenesis of implanted tumors were examined by immunostaining for platelet-endothelial cell adhesion molecule-1 and lymphatic vascular endothelial hyaluronan receptor-1 ***,we found HA promotes tumor lymphangiogenesis and the occurrence of intratumoral lymphaticvessels,but has little effect on tumor ***,HA also promotes intralymphatic tumorgrowth,although it is not sufficient to potentiate lymphatic *** results suggest that HA,which is elevated in most malignant tumor stroma,may also play a role in tumor progression by promotinglymphangiogenesis.

关键词： hyaluronan tumor lymphangiogenesis intralymphatic tumor growth

Loss of Posterior Silk Gland Transcription Specificity of Fibroin Light Chain Promoter due to Absence of 41 bp Sequence Containing Possible Inhibitor Binding Sites

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Acta Biochimica et Biophysica Sinica 2005年第12期37卷 819-825页

作者： Ting-Qing GUO~(1,2) Jian-Yang WANG~(1,2) Sheng-Peng WANG~(1,2.3) Xiu-Yang GUO~(1,2) Ke-Wei HUANG3~3 Jun-Ting HUANG~3 Chang-De LU~11 Institute of Biochemistry and Cell Biology,shanghai Institutes for Biological sciences.chinese academy of sciences.shanghai 200031.china 2 graduate school of the chinese academy of sciences.shanghai 200031.china 3 Serictdtu Research Institute.chinese academy of Agricultural sciences,Zhenjiang 212018.china Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China graduate school of the chinese academy of sciencesshanghai 200031 China Sericultural Research Institute Chinese Academy of Agricultural SciencesZhenjiang 212018 China

The gene encoding fibroin light chain protein (FibL) is specifically expressed in the posteriorsilk gland of silkworm and repressed in other *** binding sites of several transcription factorsrevolved in the silk gland transcription specificity of fibl promoter have been recognized,including SGFB,PSGF and *** we report the leak expression of the enhanced green fluorescent protein (EGFP)reporter gene in tissues other than the posterior silk gland in vivo when under the control of a shortened fiblpromoter with deletion of the 5′terminal 41 bp sequence,which is located at -650 nt to -610 nt upstream *** transcription starting *** of silk gland specificity of the promoters was performed by observationof green fluorescence in tissues of silkworm larvae following inter-haemocoelic injection of recombinantAutographa californica multiple nuclear polyhedrosis virus carrying the EGFP reporter gene controlled bydifferent lengths of fibl *** results indicated that availability of the binding sites of several knownfactors,including SGFB,PSGF and BMFA,is not sufficient for intact silk gland transcription specificity offibl promoter,and there are possible inhibitor binding sites in the 41 bp sequence (-650 nt to -610 nt)upstream of the transcription starting site which may be required to repress the activity of fibl promoter inother tissues.

关键词： promoter specificity fibroin light chain silkworm Bombyx mori recombinant AcMNPV

Effect of leukemia inhibitory factor on embryonic stem cell differentiation:implications for supporting neuronal differentiation

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Acta Pharmacologica Sinica 2006年第1期27卷 80-90页

作者： Zhao HE~(2,3) Jing-jing LI~(2,3) Chang-hong ZHEN~(3,4) Lin-ying FENG~3 Xiao-yan DING~(2,5)2 Laboratory of Molecular and Cell Biology and Laboratory of Stem Cell Biology,Institute of Biochemistry and Cell Biology:shanghai Institutes for Biological sciences,chinese academy of sciences,shanghai 200031,china 3 shanghai Institute of Materia Medica.chinese academy of sciences.shanghai 201203,china 4 graduate school of the chinese academy of sciences,shanghai 200031,china. Laboratory of Molecular and Cell Biology and Laboratory of Stem Cell Biology Institute of Biochemistry and Cell Biology shanghai Institutes for Biological sciences Chinese Academy of Sciences Shanghai China shanghai Institute of Materia Medica Chinese Academy of Sciences Shanghai China graduate school of the chinese academy of sciences Shanghai China

Aim:Leukemia inhibitory factor (LIF),a pleiotropic cytokine,has been used ex-tensively in the maintenance of mouse embryonic stem cell *** thiscurrent work,we examined the effect of the LIF signaling pathway in embryonicstem (ES) cell differentiation to a neural ***:In the presence of LIF(1000U/mL),the production of neuronal cells derived from embryoid bodies (EB)was tested under various culture *** of the LIF pathway wasexamined with specific *** effects of cell apoptosis and proliferationon neural differentiation were *** cell differentiation into three-germlayers was ***:Under various culture conditions,neuronal differ-entiation was increased in the presence of *** the LIF-activated STAT3signaling pathway with specific inhibitors abolished the neuronal differentiationof ES cells,whereas inhibition of the LIF-activated MEK signaling pathway im-paired the differentiation of ES cells toward a glial *** suppressed cellapoptosis and promoted cell proliferation during ES cell *** inhib-ited the differentiation of ES cells to both mesoderm and extraembryonic endo-derm fates,but enhanced the determination of neural ***:These results suggest that LIF plays a positive role during the differentiation ofES cells into neuronal cells.

关键词： leukemia inhibitory factor embryonic stem cells neural fate embryoid bodies

Functional Analysis of Multiple Transcription Factor Sites in a Regulatory Element of Human ε-Globin Gene

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 673-680页

作者： Chun-Hui HOU, Jian HUANG, and Ruo-Lan QIAN* State Key Laboratory of Molecular Biology, 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 State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of SciencesShanghai 200031 China

The developmental control of the human ε-globin gene expression is mediated by transcrip- tional regulatory elements in the 5’ flanking DNA of this gene. A previously identified negative regulatory element (–3028 to –2902 bp, termed ε-NRAII) was analyzed and one putative NF-κB site and two GATA sites locate at –3004 bp, –2975 bp and –2948 bp were characterized. Electrophoresis mobility shift assay (EMSA) showed that the putative NF-κB site was specifically bound by nuclear proteins of K562 cells. Data obtained from transient transfection showed that the expression of reporter gene could be upregulated about 50% or 100% respectively when ε-NRAII was inserted upstream of the SV40 promoter or ε-globin gene proximal promoter (–177 bp to +1 bp), suggesting that ε-NRAII might not be a classic silencer. Mutation in the putative NF-κB site or in the GATA site (at –2975 bp) slightly reduced the expression of reporter gene driven by SV40 promoter or ε-globin gene proximal promoter. However, the mutation of GATA site at –2948 bp remarkably reduced the reporter gene activity driven by SV40 promoter, but not by ε-globin gene proximal promoter. Further mutation analysis showed that the negative effect of mutation in GATA site at –2948 bp on SV40 promoter was not affected by the mutation of the putative NF-κB site, whereas it could be abolished by the mutation of GATA site at –2975 bp. Furthermore, the mutation of both GATA sites could synergistically reduce the reporter gene activity driven by ε-globin gene proximal promoter. Those results suggested that ε- NRAII might function differently on the SV40 promoter and ε-globin gene proximal promoter.

关键词： ε-globin gene regulatory element NF-κB GATA