细胞生物学授课教案

1(Cellbiology)211925Wilson“”12DNAmRNArRNAtRNArRNA2DNADNA345367358941.102.3.1.DNA――2.3.4.(signaltransduction)(sellapoptosis)(genomeandpost-genomicanalysis)(cellcyclecontrol)(sellapoptosis)(cellsenescence)(signaltransduction)DNA(DNAdamageandrepair)51920205060DNA11665RobertHooke40140cellar1674A.V.Leeeuwen6MalpighiGrew21838M.J.Schleiden1839M.J.Schwann“”“celltheory”.19“”1.2.“”3.18391839185918663(1)1840(Pukinje).(VonMohl)“”1861MaxSchultze1880Hanstein“”7cell(2)181RemakFlemmingStrasburgermitosis,1883VanBeneden1886Strasburger(3)1883VanBenedenBoveri18841894AltmannGolgi41)1876O.Hertwig1883VanBeneden1888Strasburger1893Overton1900341905WilsonWeissmanBorveriSutton1910Morgan2)1909819433)1924FeulgenFeulgenDNA1940BrahcetDNARNA19361940CaspersonDNARNA2051950195019609804810142×101210121g2.5-3.0200600“”“”10“”1CHONPSCAKFENACLMG112DNARNADNARNA341virusDNARNAviroidRNAprion2“”12“”3DNARNA4DNADNADNARNARNARNADNARNA5capsid“”nucleocapsid620DNARNA1322“”DNAvirion→=142-1DNADNARNARNARNARNADNARNA31.→2.→3.→→→1.2.DNAcellularoncogene(viraloncogene)3.DNARNA15“”mRNA:1.prokaryoticcell2.eucaryoticcell3.prokaryote4.eukaryote1.DNA2.3.0.210µm1.archaeobacteriaarchaeon2.eubacteria16――(mycoplast)12DNA3MRNA700450.1-0.3um6789DNA10“”DNARNA――11)2)Anucleoid,DNA17BC2DNADNA1)FeulgenDNA2)1µm312001400µmDNA20003000DNA3)DNAgenomerepliconDNADNA4)DNARNA31)182)(mesosome)3)208090%105%4)5)19flagellin4500050000mRNA570S50S30S23SrRNA5SrRNA3016SRNA206DNADNAplasmidDNA2200DNADNA7DNA8DNADNADNA20CyanobacteriaO2DNA10um70um1.DNADNADNADNA“”235nm3αβ14α→→α21aab34451.2.223.4.22DNADNA2DNA70S50S30S80S60S40Saa,bDNADNADNADNA2323DNADNA12DNA1n2nn“”“”DNADNA5DNADNA24DNA100DNA2-4DNADNADNADNA70S5580S7084605SrRNA251.2.DNARNA3.810nm26DNARNADNARNADNArRNArRNA80S1525nmmRNA2725/µm0.020.10.3122030(1050)281.2.3.(1)(2)29(3)CO2“”DNARNA0.2mm0.2µm0.2nm301cm1mm100um10um1um100nm10nm1nm0.1nmlightmicroscopy123λαND0.61λ/N.sinα/2α140N1λ450nmD292nm0.3umN1.5D0.2um0.2um.fluorescencemicroscopy31greenfluorescentprotein,GFP――phade-contrastmicroscope“”“”differential-interferencemicroscopevideo-enhancemicroscopy32electronmicroscope0.1nm31200nm100nm200nm0.1nm0.01-0.9nm1.33×10-5-1.33×10-3Pa20.2mm0.2um0.2mm/0.2um10000.2nm1063a)b)33c)d)1um14050nm20um342.negativestaining1.5nm3“”“”4Xstructuralbiology355scanningelectronmicroscope,SEM“”“”scanningtunnelingmicroscope,STM100nm10.10.2nm0.001nm23DNARNAatomicforcemicroscopedifferentialcentrifugation,――36S1)FeulgenDNA2)PAS3)immunofouorescencetechnique37insituhybridizationDNARANAgBrAgClpulse-chase3333β32Pβ14.214Cβ10um57603Hβ0.8um12.335Sβ10um-1mm87131Iβ10um-1mm8DN3HTdR3HRNA3HU35S3H14C38microspectrophotometryflowcytometryDNARNA1/ms1000/h39primaryculturecellsubculturecell110singlelayercellcellstrain1040104050cellline50HeLaBHK21CHOfibroblast(epitheliallikecell)1――2――protoplastcell-freesystemDNARNAcellengineeringcellfusionPEG2080electronfusionmethod41100200V/cmhomokaryonheterokaryonsynkaryon(monoclonalantibody)BcytochalasinBkaryoplastcytoplastPEGreconstitutedcell42micromanipulationmicroinjectionmRNAfunctionalgenomicsproteomics(bioinformatics)DNA43cellmembraneplasmamembrane(biomembrane)1924E.GorterandF.GrendelDavsonandDanielli“”1959J.D.RobertsonunitmemrbranemodelX1972S.J.SingerandG.Nicolsonfluidmosaicmodel1)2)“”“”“”(lipidraftsmodel),“”441)2)3)109um25×106()350%4-11/35%451)108cm2/s2um2)3)4)liposome()peripheralproteinsextrinsicproteinsintergralproteinstransmembraneproteins1)2)3)1)20α2)αα463)porins1012βdetergent1)SDS2)TritonX-10010min40minpatchingcapping47fluorescencerecoveryafterphotobleaching,FRAPextrocytoplasmicsurface,ESprotoplasmicsurface,PSextrocytoplasmicface,EFprotoplasmicface,PFES14823456membraneassociatedcytoskeletonO2CO2spectrinankyrin34.1actinglycophorin49Cαβ100nm5nm200nm10491.occludingjunctionstightjunction2.anchoringjunctions1)desmosome(hemidesmosome)2)adhesionbelt(focaladhesion)3)communicatingjunctionsgapjunctionchemicalsynapseplasmodesmata.1.1occluding60×10342Clauding4152.501)attachmentproteins,2)basementmembranebeltdesmosome.513.1.23nm4142.(connexon)6connexin1.5nm204α3.zz1×103z1DNADNADNADNA52DNAcAMPCa2+2synapsedlectronicjunction34PHCa2+→→→→cAMP↑→cAMP↑→→→↑→cAMP→53movementproteins4-254—cadherins43416417integrins4443Ca2+Mg2+ENPPNα1β22055α6β4syndecans44(desmogleinsdesmocollins)DesmoplakinsplakoglobinEvinculinα6β4Desmoplakin-likeTalin,vinculin,α-actinin156E72075050%60%N54Ca2+2selectinCa2+lectinPplateletE(endothelial)L(leukocyte)3CAMIg-superfamilyCAMCa2+NCAMs(nerve-celladhesionmolecules).NCAM4αβ16α8β2257RGDcellcoatglycocalyxextracellularmatrix4191.collagen30%204545[α1]2[α]300nm67nm[α1]3300nm67nm58[α1]3300nm67nm[α1]2[α2]390nmC[α1]2[α2][α3]3390nmN[α1]2[α2][α3]150nmNC100nmα1α2α32300nm1.5nm4201/31/43.procollagenpreprocollage459glycosaminoglycan42146haaluronicacidproteoglycancoreprotein90%95%linkerprotein4µmFGFTGF60lamininAB1B2αβγ8α1α2α3β1β2β3γ1γ273422Arg-Gly-Asp(R-G-D)nidogenfibronectin4.5-9.5%220×103250×103C20RGD12elastin6183012Lysβ1→4microfibrilCa2+extensin30015%Ser-Hyp-Hyp-Hyplignin62passivetransportsimplediffusionPKDPKD/ttfacilitateddiffusion107cm/s63102cm/s10512VmaxKM3KM4membrane