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检索条件"任意字段=中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议"

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A lentiviral vector visualizing the germ cell specication in vitrounder the control of Figla promotor

A lentiviral vector visualizing the germ cell specication in...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Zhili Chu Bowen Niu Na Li Weishuai Liu Yue Hu Ping Yu Jian Li ChongyangWu Xinrong Yan Anmin Lei Jinlian Hua College of Veterinary Medicine Shaanxi Centre of Stem Cells Engineering & Technology Northwest A&F University Department of Patholohy Yangling Demonstration Hospital College of Life Science Northwest University

Premature Ovarian Failure(POF)is affecting more and more women,which is the loss offunction of the ovaries before *** elucidate the underlying mechanisms of the oogenesisis of importance to understand the causes of impaired fertility and ***,Oogenesis is a complex process in ***,building an oogenesis visualizing system isbeneficial for the studyof *** this study,we found that Figla is specifically expressedin female mice *** we constructed a lentiviral vector(pTRIP-Figla-EGFPpuro)under the control of Figla promoter,which drived enhanced green fluorescent protein(EGFP)as a indicator,and used the lentiviral vector transduction the ovarian cells andinduced germ cells derived from hUC-MSCs,and the results showed that the lentiviral vectorwe constructed was able to specifically express green fluorescent protein(GFP)in theovarian oocyte and induced oocyte-like cells derived from human umbilical cord mesenchymalstem cells(hUC-MSCs),which was Figla positive *** results suggest thatpTRIP-Figla-EGFP vector provides a new system to study the role of Figla in oogenesis,andan approach to study the development and the differentiation of germ cells derived fromstem cells.

关键词： Figla Oogenesis Lentiviral vector Induced differentiation human umbilicalc ord mesenchymal stem cells(hUC-MSCs)

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The biosynthesis of DHA is increased in the liver of diabetic rats induced by high-fat diets and STZ, in correlation with increased activity of peroxisomal β-oxidation

The biosynthesis of DHA is increased in the liver of diabeti...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Min Yao Key Laboratory of Neural and Vascular Biology Department of Biochemistry and Molecular Biology Ministry of Education Hebei Medical University

Background: Docosahexaenoic acid(DHA) biosynthesis in the liver is the most importantsource of extra-hepatic tissue DHA. However, the levels of DHA and DHA biosynthesis inthe liver of type 2 diabetes remain ***: In the current study, we used type 2 diabetic rats induced by high-fat diets andlow-dose streptozotocin(STZ) to investigate DHA biosynthesis in the liver of type 2 *** acid profiles, the activity of peroxisomal β-oxidation and the expressions ofenzymes involved in the DHA biosynthesis pathway were assayed in the liver of *** and methods: The data showed that DHA levels were unexpectedly increased despiteof the lower expressions of key enzymes, △5- and △6-desaturases(△5D and △6D) in theendoplasmic reticulum(ER) with this effect being due to insulin resistance in the diabeticliver. The increased DHA was accompanied by an increase in the expression of straightchainacylCoA oxidase(SCOX), D-bifunctional protein(DBP) and 3-oxoacyl-CoA thiolase(THL), as well as elevated activity of peroxisomal β-oxidation and an increased capacity forDHA biosynthesis from C24:***: These findings indicate that DHA biosynthesis is increased in the livers of diabeticrats induced by high-fat diets and STZ, and that this effect is related to increasedactivity of peroxisomal β-oxidation, but not of ER desaturases.

关键词： DHA peroxisomal β-oxidation

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Structural insights into VirB-DNA complexes reveal mechanism oftranscriptional activation of virulence genes

Structural insights into VirB-DNA complexes reveal mechanism...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： xiao xiao pan gao sheng cui 医科院病原所

VirB activates transcription of virulence genesin Shigella flexneri by alleviating heat-stablenucleoid-structuring proteinmediated promoterrepression. VirB is unrelated to the conventionaltranscriptional regulators, but homologous to theplasmid partitioning proteins. We determined thecrystal structures of VirB HTH domain bound bythe cis-acting site containing the inverted repeat,revealing that the VirB-DNA complex is related toParB-ParS-like complexes, presenting an examplethat a ParB-like protein acts exclusively in transcriptional regulation. The HTH domain ofVirB docksDNA major groove and provides multiple contactsto backbone and bases, in which the only specificbase readout is mediated by R167. VirB only recognizes one half site of the inverted repeatscontainingthe most matches to the consensus for VirB *** binding of VirB induces DNA conformationalchanges and introduces a bend at an invariantA-tract segment in thecis-acting site, suggesting arole of DNA remodeling. VirB exhibits positivecooperativity in DNA binding that is contributed bythe C-terminal domain facilitating VirB oligomerization. The isolated HTH domain onlyconfers partialDNA specificity. Additional determinants for sequence specificity may reside in N- or Cterminaldomains. Collectively, our findings support and extend a previously proposed model for relieving heat-stable nucleoid-structuring protein-mediated repression by VirB

关键词： shigella H-NS VirB

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Regulation of host NF-κB signaling pathway by a M. tuberculosissecreted protein interacting with host E3 ubiquitin ligase TRIM27

Regulation of host NF-κB signaling pathway by a M. tubercul...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Cuihua Liu Jing Wang Bingxi Li Institute of Microbiology Chinese Academy of Sciences

Mycobacterium tuberculosis(Mtb) secretes a variety of effector proteins into host cellsduring the infection process. Those secreted proteins could interfere with a variety of hostsignaling pathways to promote the intracellular survival of Mtb. NF-κB signaling pathway iscritical to the regulation of host innate immune response by controlling expression of anumber of immuno-regulatory molecules(e.g., TNF and IL-1β), thus it is frequently modulatedby intracellular pathogens such as Mtb. Here, we demonstrate that the nonpathogenicM. smegmatis over expressing a secreted Mtb protein, which we namedMUIP2, exhibited an increase in bacterial load and suppression of NF-κB signaling activationin macrophages. Further investigation through yeast two-hybrid and Co-Immunoprecipitation assays revealed that MUIP2 interacts with a host E3 ubiquitin ligaseTRIM27. Dual-luciferase reporter assay data demonstrated that TRIM27 could also inhibitNF-κB signaling activation, and this inhibition could be enhanced by the addition ofMUIP2. Collectively, our data suggest that MUIP2 promotes intracellular survival of mycobacteriaby suppressing host NF-κB signaling pathway through interacting with TRIM27.

关键词： M.tuberculosis NF-κB signaling ubiquitin ligase

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Multiple copies of a linear donor fragment released in situ from avector improve the efficiency of zinc-finger nuclease-mediated genomeediting

Multiple copies of a linear donor fragment released in situ ...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Weifeng Zhang Dan Xiao Dongyang Wang Xiaojing Zheng Haibin Xia Shaanxi Normal University

Homologous recombination(HR) is a strategy for genetic correction. The efficiency of HRcan be increased by creating a targeted double-strand break(DSB) via zinc-finger nucleases(ZFNs) and /or by introducing linear donor DNA intracellularly. Some studies havesuggested that increased copy numbers of linear donor DNA may further improve HR ***, the introduction of multiple copies of a linear donor fragment remains achallenge, particularly in cell types with low transfection efficiency. In this study, we developeda vector that carries tandem repeats of a donor fragment, with each repeatflanked by ZFN target sequence fragments(TSFs). The cleavage of the flanking TSF sequenceby ZFN would lead to the release of multiple linear fragment. We demonstratedthat this novel vector carrying 5 copies of a linearizable donor fragment, when cotransfectedwith a ZFN-expressing vector in 293 cells, showed improved HR efficiencyabout 30 times, as compared with vector carry non-linearizable donor. For the applicationof gene therapy, we then introduced this system into an adenoviral vector, which also revealedmarkedly improved ZFN-mediated HR efficiency in cells. The novel strategie spresented here have the potential to promote the application of ZFNs in both basic researchand disease therapy.

关键词： zinc-finger nuclease homologous recombination adenoviral vector site-specifici ntegration genome editing

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Modulation of gene expression following cellular senescenceinhibitedgene silencing in HEK293 cells

Modulation of gene expression following cellular senescencei...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Liwei Ma Wenting Zhao Feng Zhu Tanjun Tong Peking University

Cellular senescence-inhibited gene（CSIG）, also named ribosomal1 domain-containing 1（RSL1D1）, is implicated in various process including cell cycle regulation, cellular senescence,apoptosis, and tumor metastasis. However, little is known about the regulatorymechanism underlying its function. To screen important targets and signaling pathwaysmodulated by CSIG, we compared the gene expression profiles in CSIG-silencing and thecontrol HEK293 cells using Affymetrix microarray Human Genome U133 Plus 2.0 GeneChips.A total of 121 genes analyzed had known functions and displayed statistically significantexpression changes, with 57 up-regulated and 64 down-regulated respectively. Thesegenes were involved in a broad array of biological processes, mainly in cell cycle, signaltransduction, transcriptional regulation, mRNA transport, and cell adhesion. The differentialexpression of genes such as ZNF616, KPNA5, MAP3K3 were further validated by realtimePCR and western blot analysis. In addition, we observed the correlated expressionpattern between ESCO1, KPNA5, MAP3K3 and CSIG during cell cycle and senescence progression,suggesting the important path ways by which CSIG regulates cell cycle and senescence.

关键词： CSIG/RSL 1D1 senescence

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Selection of small molecules targeting Dclk1 of tumor stem cellsby phage display and computer aided virtual screening

Selection of small molecules targeting Dclk1 of tumor stem c...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者： Yijie Zhou Bowen Tan Yinsong Dong Canquan Mao Southwest Jiaotong University

Targeted therapy is a promising way for cancer based drug development. Among variouscancer associated targets, tumor stem cells(TSCs) hold great potential as recent discoveryof their important role in cancer development and resistant to therapy. Doublecortin-likekinase1(Dclk1) is a serine-threonine kinase which has been found as a marker in tumorstem cells. In our study, we desired to find anticancer drugs targeting DclK1 of TSCs basedon phage display and computer aided virtual screening for cancer *** the basis of bioinformatics analysis of Dclk1 family, we found a divergent region whichdiscriminate from other Dclk members. We synthesized the peptides and random phagedisplay peptide library was used to screen the binding peptides for Dclk1. After threerounds of selection, the phages were enriched, four peptides were obtained and furtherdetermination of the peptides was still in progress. On the other hand, 100,000 small moleculespicked up randomly from ZINC database were virtually screened against the NterminalDCX domain of human Dclk1, the top 10 small molecules were reserved and analyzedfor their interactions with the receptor, which also provided a basis for the developmentof small chemical molecule drugs targeting Dclk1.

关键词： Dclk1 peptide inhibitors virtual screening bioinformatic phage display

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杜仲雌雄株转录组测序数据组装及基因功能注释

杜仲雌雄株转录组测序数据组装及基因功能注释

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：赵德刚李岩吕立堂赵丹董璇 Yi Li 贵州大学生命科学学院/农业生物工程研究院山地植物保护与种质创新省部共建教育部重点实验室贵州大学生命科学学院/农业生物工程研究院山地植物资源保护与种质创新省部共建教育部重点实验室贵州大学生命科学学院/农业生物工程研究院山地植物资源保护与种质创新省部共建教育部重点实验室贵州大学生命科学研究院/农业生物工程研究院山地植物资源保护与种质创新省部共建教育部重点实验室 Department of Plant Science University of ConnecticutStorrsCT06269USA

杜仲(Eucommia ulmoides Oliv.)是具有重要开发价值的药用植物和胶原植物,建立转录组数据库对分离关键功能基因、阐明基因表达特性具有重要意义。本研究在杜仲幼果发育时期对雌雄株幼果、叶片、树皮的转录组进行测序,构建了转录组数据库... 详细信息

杜仲(Eucommia ulmoides Oliv.)是具有重要开发价值的药用植物和胶原植物,建立转录组数据库对分离关键功能基因、阐明基因表达特性具有重要意义。本研究在杜仲幼果发育时期对雌雄株幼果、叶片、树皮的转录组进行测序,构建了转录组数据库,获得雌株51,574,000条、雄株52,430,502条Clean Reads数据,分别包含总长度为4,641,660,000nt和4,718,745,180nt序列数据信息;经拼接组装,获得雌株基因信息长达69,461,730nt的423,339个Contig片段,获得雄株基因信息长达94,814,201nt的542,383个Contig片段;对Scaffold进一步拼接,分别获得平均长度为229nt的雌株159,434个Unigene片段和平均长度为231nt的雄株257,298个Unigene片段;共有48761个表达序列标签(EST),以Unigene和NR数据库比对,杜仲雌雄株转录组基因序列与葡萄的同源性最高,为33.8%,其次是蓖麻(11.4%)和杨树(11.2%),与拟南芥的同源性很低,才2.3%;以Unigene和COG数据库比对,可将杜仲转录组Unigene的功能分为25类,根据GO功能可分为生物过程、细胞组分和分子功能3大类55分支;与KEGG数据库比较,杜仲雌雄株转录组数据含有127类代谢通路的基因,包括多种氨基酸生物合成、脂肪酸代谢、糖代谢,尤其是次生物质代谢途径较多,如萜类化合物生物合成、黄酮与黄酮醇生物合成、异喹啉生物碱生物合成、花青素生物合成等。

关键词：杜仲转录组功能基因

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Oxacyododecindion a novel HIF-1 inhibitor

Oxacyododecindion a novel HIF-1 inhibitor

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：魏晓艳北京师范大学生命科学学院生物化学与分子生物学研究所

Nuclear transcription factor one(HIF-1) plays an eminent role in the process ofcellular internal oxygen sensing system, increased levels of HIF-1 can increase the tumormetastasis therapeutic resistance and poor prognosis. In this study we discovered that oxacyododecindiona fungi-derived macro cyclic lactone have anti-tumor effects and allthese functions were achieved through specifically decrease the level of HIF-1α in multilevel with out influencing HIF-2α.RT-PCR assay suggest that oxacyclododecindion can specificallydecrease the mRNA level of HIF-1 α increase the mRNA level of degradation relativeproteins including VHL, PHD3, P53, MDM2,Western blot suggest that oxacyclododecindion distinctively decrease the synthesis of HIF-1α mainly through the PI3K-AKTmTORand promote the degradation of HIF-1αthrough up regulate the protein level ofVHL,PHD3. Reporter assay combined with plasmid transfection assay showed that oxacyclododecindion can decrease the HIF-1 activity in vivo including the secret of cytokines suchas VEGF and GLUT1. Cell transfection combined with western blot suggest that oxacyclododecindion has the ability to inhibit HIF-1α from into nuclear and this is in relate withMEK- ERK signaling pathway, wound closure and HUVEC tube formation assay suggestthat the invasion metastasis and angiogenesis ability of HUVEC was depressed *** and flow cytometry combined with cell transfection proved Oxacyododecindion can impede the cell cycle in a concentration dependent manner and increase the apoptosisrate of HepG2 cells mainly through the caspase3 pathway, the cell toxicity of oxacyclododecindionin HepG2 and HUVEC cell lines is lower than0.27 nM. These data suggest that oxacyclododecindion have great potential to be developed as high efficiency and low toxicityantitumor drug.

关键词： HIF-1 Oxacyododecindion

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Denaturant effect on free energy of protein unfolding process

Denaturant effect on free energy of protein unfolding proces...

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中国生物化学与分子生物学会第十一次会员代表大会暨2014年全国学术会议

作者：李茂东刘志荣北京大学定量生物学中心

Protein denaturation refers to its specific spatial conformation change according to thecertain physical or chemical *** of the research in the past is carried out on theensemble *** was found that the free energy change was linear with the concentrationof *** ensemble describes average properties of the system,but the probabilisticinformation of different states in the system is *** single molecule methodsmake it possible to work on these *** are two main models for protein denaturation,one is Transfer model,another is Binding *** analyzed the free energychange in the process of protein denaturation and the universality of the two *** the transfer model,it was found that the protein residue interactions energy differenceεdepends linearly on the denaturant concentration,and that the free energy of denatured-state collapse has a linear dependence on denaturant *** we foundthat the collapsed state was far away from the equilibrium state,which could only providea reference state for native state but not an intermediate *** that the coil-to-globuletransition of the denatured state didn’t appear in protein folding,suggesting that conformationalentropy of the denatured state was not so important as *** addition,we found out the neglected collapsed-to-native free energy could describe the specificinteraction in folding,which was independent of the denaturant concentration but linearlydependent on residue *** the binding model,it was assumed that there are identical independent binding siteswith an effective association constant,which could not satisfy the linearity between thefree energy change and the denaturant *** achieve a linear relationship,itwas found that an abnormal dependence of activity on the denaturant concentration hasto been *** conclusion,Transfer model is a good mechanism for protein denaturation,while Bindingmodel remains to be optimized.

关键词： Protein denaturation transfer model binding model

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