简介:Witharapidincreaseinvolumeofve-hiclesintheworld,thepolutioncausedbytheautomobileexhaustedgashasbe-comemoreandmoreserious.Thisproblemhasbroughttoourgovernment’satten-tion.InBeijing,regulationsregardingthesupervisionoftheexhausthavebeenissued.AttheFifthNationalMeetingonRECatalyticpurificationofAutomobileEx-haustheldinBeijing1989,thepartici-pantsfrominstitutes,universities,andfactoriesalloverthecountryreportedanddiscussedthemethodsofpurifica-
简介:Separationandpurificationofhumanchorionicgonadotropion(HCG)intheurinesampleofearlypregnantwomenbyD3520resinadsorptionchromatographyisreported.ThecrudeproductobtainedbyDEAE-Cellulose23andDEAE-SephadexA50columnchromatographyshowedahighactivityofHCG.FurtherpurificationofthesamplebygelfiltrationchromatographyonaSephadexG75columngivesafinalpreparationof6000-6500IU/mg.ThepreparationmeetstherequirementsofthepyrogntestinChineseLawofPharmacopeia.
简介:AhepaticstimulatorsubstancefromSharkliver(sHSS)waspurifiedandcharacterized.Thepurificationprocedureincludedhomogenization,centrifugation,ultrafiltration,DEAE-sephadexA25chromatographyandFPLCmonoQchromatography.Bytheprocedure,thestimulatoryspecificactivityofsHSScouldbeincreasedto9000IU/mg,andwas750timesmorethanthecrudeextract.ThepurityofthefinalpurifiedfractionFTowhichretainedthemaximalactivityofsHSSwascheckedbypolyacrylamidegelelectrophoresisinthepresenceofsodiumdodecylsulfate(SDS-PAGE)andHighPerformanceGelFiltrationrespectively,andshowedonlyonebandoronepeak.Itsmolecularweightwasmeasuredtobe16973Dabymassspectrometry.ItsN-terminalsequencewasdeterminedtobeN-Met-Arg-Thr-Gln-Glu-His-Thr-Lys-Ser-ValbyPE-ABD491AProteinN-TerminalSequencingMeter.ItwasstableoverawiderangeofpHandtemperature.
简介:FADD是在死亡的一个重要proapoptotic适配器导致受体的apoptosis。最近,FADD被发现了参予许多non-apoptotic过程,例如开发,房间周期前进和幸存。它的non-apoptotic活动被在C终端区域定位的丝氨酸残余的phosphorylated地位调整,从proapoptotic函数不同的域联系了DED和DD域。由于在自然FADD的表示和结晶化的困难,然而,迄今为止,所有FADD变体的分子的结构没包含C终端区域。阐明C终端区域的结构功能关系,我们需要获得FADD变体那个包含的C终端区域。在这研究,包含DD领域的鼠标FADD(80-205)和C终端区域,指定了为C-FADD,在E被表示。有在N终点的他的标签的coli并且由Ni2+亲密关系层析净化了。净化的蛋白质在glutaraldehydecross-linking分析作为同质的单体存在并且在CD(圆形的二色性)展出了一个典型螺旋系列试金。在vitro他的标签,下拉试金证明净化的C-FADD拥有了CK为它的non-apoptotic功能重要的我有约束力的活动。
简介:Adiatomwaspurifiedwithcolonyselectionandcontinuousdilutionmethods.ItwasidentifiedtoCylindrothecaclosteriumaccordingtoitsmorphologicalcharacteristicsandrbcLand18srRNAgenesequences.Thealgawasnotsensitivetoampicillinandneomycin,butsensitivetochloramphenicolwhichinhibiteditsgrowthatconcentrationsrangingfrom50to150μgmL-1.Thepurifiedalgawaseasytocultureanditsspecificgrowthratewas0.207±0.002(d-1).Itwasresistanttopollutionandcouldbeharvestedinaneasyway.Itwasrelativelyhighinlipidcontent(20.08%±0.67%ofdryweight)andthecombinedamountofits16:0and16:1(n-7),themostsuitableresourceofbiodiesel,wasashighas64%ofthetotalfattyacids,whiletheamountofpolyunsaturatedfattyacidsreached19.96%–20%ofthetotalfattyacids.ThusthepurifiedC.closteriumcanbeculturedasabiodieselproduceroranutritionsupplementproducer.
简介:Theeffectofresinstructureondesalinationofligninsolutionwasinvestigated,theoptimalstructureofresinisasfollows;crosslinkingdegreeas4%,ratioofcationogentoanionogenisnear1.withsuchresinthedesalinationofligninwasproducedverywellbecausetheresinhasbothmoleculesievingandionretardationproperties.Thesulfonationdegressofligninandtotalsaltcontentofligninsolutionweredeterminredwithionexchangetechnique,therelativeerrorlessthan1%.Thesaltcontentofsmallmoleculeintheligninsolutionwascalculatedfromsulfonationdegreeofligninandtotalsalt.Amonggelandmacroporousresinsthebestseparationofligninfromreducingsugarwasachievedwithinterpenetratingsulfonatedresin2×1.5×1.Theseparationofligninwithinterpenetratingresinwascarriedoutsimultaneouslywithfractionationoflignin,theeffectoffractionationwithmacroporoussulfonatedresinisbetterthanthatwithinterpenetratingresin,buttheformerhasadefinitesorptionofligninwhichdecreasedtherecoveryoflignin.
简介:Objective:Toachieveanoptimizedmethodforsolubleexpressionofhumancarboxylesterase1(hCE-1)inescherichiacoilandpurificationbyNi2+-NTAagaroseaffinitychromatography,togetimprovedproteinyieldandpurityforfurtherdevelopmentofhepatocellularcarcinoma(HCC)diagnosisELISAkits.Methods:ThebestantigenepitopesofhCE1werepredictedbycomparingsecondarystructure,flexibleregions,hydrophilicity,antigenicindexsurfaceprobabilityofresidues.Afterwards,pET-42a(+)withaHis-tagandaGST-tagwasappliedtoformrecombinantplasmidpET-42a(+)/hCE1,whichfacilitatedpurificationwhenusingNi2+-NTAagaroseaffinitychromatography.ProteinqualitywasmeasuredbySDS-PAGEandBCAproteinassay.Western-blotidentificationwasalsoperformedtoensurethecorrectexpressionofhCE1protein.Results:Theresiduesfrom500to567nearC-terminalofhCE1proteinwereconsideredthebestepitopeswhichexhibitedhighhydrophilicityandhighsurfaceprobabilityandrelativelyflexiblesecondarystructureandlowhomologycomparedwithhCE2andhCE3.His-hCE1500-567fusionproteinwasachievedbyIPTG-inductedexpressionwithanexpectedmassof42kDa.Afterpurification,thefinalproductwasspeciallyidentified,whichreachedover95%purityandmorethan10mg/Lofmicrobialculture.InWesternblot,thepurifiedfusionproteinwasrecognizedbyanti-hCE1monoclonalantibody,alongwithprevioussequencingvalidation,whichdemonstratedthecorrectpreparationofsolublehCE1protein.Conclusion:ThisisanefficaciousandaffordablestrategytogeneratefusionhCE1ofhighqualityinEcoli,whichfacilitatespreparationofhCE1monoclonalantibodyandfurtherHCCdiagnosisresearch.
简介:Abstract:Aphysiologicalactivefactor-r-oryzanolinricebranwasstudied.Ther-oryzanolwasseparatedandidentifiedbythinlayerchromatograghy(TLC),reverse-phaseHPLC,semipreparativeHPLCandelectrosprayionization/massspectrometry(ESI/MS).Itsindividualcomponents,cycloartenolferulateand24-methylenecycloartenolferulatewereobtained.Aneffectivemethodtopurifyandseparatether-oryzanolisprovided.Whichisusefulforfurtherresearchonthefuctionalclaracteristiesofr-oryzanolandforthedevelopmentofnewricebranproducts.
简介:Weproposeaschemetopurifyentanglementoftwoatomsfromnot-too-impureentangledstatesbycheckingtheparityofthetwoatomsthroughthecavityinput-outputprocess.Astheparitycheckismadebymeasurementonsingle-photonpolarization,whichwouldnotaffecttheentanglementofthetwoatoms,ourschemehasthesuccessfulprobabilitydoubleofthatinapreviouswell-knownschemewithlinearopticalelements[Nature(London)410(2001)1067],andisinsensitivetothephotonlossandthedetectioninefficiency.Experimentalfeasibilityofourschemewithcurrenttechnologyisdiscussed.
简介:ThepurificationprocessoftotalflavonesofginkgoleavesbyresinHZ-841fromethanolextractwasstudied.First,thetotalflavonewasextractedfromthedefattedpowderofginkgoL.bilobaleaves.Effectsofsolventsandoperationconditionswereexaminedtogetarelativehighyieldandpurityinthisstep.ThecrudeextractwasfurtherpurifiedbyresinHZ-841.Bothadsorptionandelutionprocesswerestudiedtoobtainanoptimizedconditions,i.e.,pH,flowrate,concentration.Ayellowpowderwasobtained,ofwhich37.3%wasflavones,obviouslyhigherthanthebasicinternationalstandardof24%.
简介:Thepowderedactivatedcarbontreatment(PACT)processhasbeenwidelyusedinmanyindustrialfields,however,veryfewPACTprocessesarebuiltforpetrochemicalwastewatertreatmentinChina.AnindustrialPACTunitlaunchedinapetrochemicalplantwasintroducedandevaluatedfromboththepracticeandmechanismstudy.Practically,thePACTprocessshowedexcellentcapabilityinpollutantsremoval,shockresistance,toxicitytolerance,andtheCODandammonium-NineffluentofPACTunitassistedbyPACwasequalto15.5mg/Land0.7mg/LlowerthanthatwithoutPACaddition,respectively.ThewetoxidationregenerationunitwasquiteefficientinsupplyingregeneratedPAC,and,however,thehardcalciumsulphatescaleandthehighpollutantconcentrationsolutionneededtobecarefullycontrolled.Moreover,althoughthecarbonbalanceshowedthattheadsorptioncapabilityofregeneratedPACwasnegligible,thebiologicaltestsprovedthattheregeneratedPACincreasedmicrobeactivityupto17%morethanpureactivatedsludgesystem,whichwasalmostcompatiblewiththefreshactivatedcarbon.
简介:
简介:Sumoylationisanimportantproteinmodificationdiscoveredrecently.SUMO(smallubiquitin-relatedmodifier)pathwayregulatestheproteinstabilityandtranscriptionalactivitywitha12-kDasmallmolecularprotein,SUMO,ligatedtothetargetprotein.ThepurificationofSUMOproteinsisakeysteptorevealtheirfunction.ThepurposeofthisstudywastoconstructtherecombinantSUMO1geneclonedtoapGEX-4T-1vectortoexpressandpurifytheSUMO1-GSTfusionproteininEscherichiacoli.First,thefulllengthDNAsequenceofSUMO1genewasamplifiedbyPCRandwasligatedtopMD18-Tvector.ThentheSUMO1genewassubclonedtopGEX-4T-1prokaryoticexpressionvectorbetweenBamHIandXhoIsites,andtransformedinEscherichiacoliDH5αcells.Therightcolonieswereidentifiedbyrestrictiveenzymedigestionandsequencing.ThecorrectrebombinantplasmidofpGEX-4T-1-SUMO1wastransformedinEscherichiacoliBL21cellsandtheninducedbyIPTG(isopropyl-β-D-1-thiogalacto-pyranoside)toexpresstheSUMO1-GSTfusionprotein.ThehighlypurifiedSUMO1-GST(glutathioneS-transferase)fusionproteinwasobtainedbyaffinitychromatography.Finally,thepropertiesofSUMO1-GSTfusionproteinwereconfirmedbyCoomassiebrilliantbluestrainandWesternblotanalysis.TherecombinantplasmidofpGEX-4T-1-SUMO1wassuccessfullyconstructed,andSUMO1-GSTfusionproteinsweresuccessfullyexpressed.
简介:AdsorptionresinAASwasusedfortheseparationandpurificationofmilupeinan(M-13)fromitsmotherliquor.ThegoodadsorptionpropertyofAASadsorptionresinwasfoundformilupeinan(M-13)comparedwiththeDiaionCHP20p.Whenacetoneandwater(2:1involume)wereusedasdesorptionreagents,itwasfoundthat3timesofbedvolumeofthedesorptionreagentswouldmaketheAASresinwhichwassaturatelyadsorptedM-13desorptioncompletely.Alsoamacroporousanionexchangeresin(Poly-4-vinylpyridinetype)D-280wasusedforthedecolourizationfromM-13solution.Goodresultswasgivenwhenthesr=1~2.
简介:PhycoerythrinandphycocyaninwerepurifiedfromPorphyrayezoensisUedawiththeirbioactivitydeterminedinthisstudy.Continuousprecipitationwithammoniumsulfateatdifferentconcentrations(10%,20%,40%and50%)increasedthepurity(A564:A280)ofphycoerythrinto1.49,3.92foldoftherawextract(0.38)andthepurity(A615:A280)ofphycocyaninto0.70,3.33foldoftherawextract(0.21).Twomoretimesofchromatographywithhydroxylapatitesfinallymadethepurityofphycoerythrinandphycocyaninreach5.50,14.47foldoftherawextract,and5.10,24.29foldoftherawextract,respectviely.Theyieldofhighpurityphycoerythrinandphycocyaninwere0.21%and0.09%ofdriedP.yezoensisblade,respectively.Thephotodynamiccytotoxicexperimentshowedthatbothphycoerythrinandphycocyanininhibitedthegrowthoflivertumorcellssignificantly.Itwasfoundthat250mgL-1purifiedphycoerythrinandphycocyanininhibitedthegrowthofhepatocellularcarcinomacells24hafterlaser-irradiationby80%and59%,respectively,and100mgL-1purifiedphycoerythrinandphycocyanininducedtheapoptosisof31.54%and32.54%ofthecells,respectively,8hafterphotodynamictherapy.OuefindingsdemonstratedthatP.yezoensiscanserveasphotosensitizer(phycoerythrinandphycocyanin)producer.