分子生物学试验技术试验报告

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7、kaielulgpilkaieclueptieclueptiethooghtiethooghtieststpyofbasccitell,eailguidesaidXIGpeaZedayiiFuaIwrkduilgllvcaeofourgras.ittpyofbasccitell,eailguidesaidXIGpeaZedayiiFuaIwrkduilgllvcaeofourgras.itt、.oaeyiidodipa.oaeyiidodipa，bldflsfyaidlheINaiplgreeauHud,plposbldflsfyaidlheINaiplgreeauHud,plposof.lg

8、all,Xa.ov,adiiov.Ii.eLais.,.potatthouof.lgall,Xa.ov,adiiov.Ii.eLais.,.potatthou，tseraidse.gthethepayspll,uHlsthepurposetseraidse.gthethepayspll,uHlsthepurposeias.oraLhaaLercisias.oraLhaaLercisato.reatato.reattothebascequroetsMa.uiuesad.IsteflotothebascequroetsMa.uiuesad.IstefloIgaseCsLiiteit(IgaseCs

9、Liiteit(PCRBuffer2.5始dNTPs2始P11始P21始Taq酶0.25cDNAtemplate2LTotle252、设立阳性对照组和阴性对照组。3、加完体系后，轻弹混匀，瞬离，上PCR仪。4、特异性反应条件为：6 6CuyeciomicIf.InaCuyeciomicIf.InaILIL-les.-les.，.,一一. I Ia aLOLO- -lea11garai.me-sspeclkprlea11garai.me-sspeclkpra.fora.for. .ulul”yrUeparyLlgtyrUeparyLlgt，.”0 0111111aton.itoiCuy,11at

10、on.itoiCuy,nanaLOLO lea11garailea11garai.eit.ofseclkpro.eit.ofseclkpro”.,.,. .L LIBoaIBoaL L-rii.L-Hi.p.m.-iaonp.-rii.L-Hi.p.m.-iaonp.espit.espitordigt.ordigt.”. .a aILILh-th-t”.Ulip.Ulip.of.i-.ts,.of.i-.ts,. .a.ing.a.ing.ion.*.*promo.alm-b.scnsl.ion.*.*promo.alm-b.scnsl.esectdi.esectdi.“on,aion,aiL

11、 L-.y-.y- -aruaru,wt.iGe.c.la,wt.iGe.c.la，se.s.por.ntspecsilse.s.por.ntspecsil.ILIL.I.IL L,.,.L L.pracbeaipracbeaiL LPromo.teworkaPromo.teworkaILILbequOi-pa.bequOi-pa.be.be. .L L.ilg.ng.ilg.ng.eh.eh.，mamatefolo-Wigspeciepr.mm-.tefolo-Wigspeciepr.mm-.ingr.ingri-.01alpa.i-.01alpa.be.t.be.t.，.payi.In.p

12、ayi.In. .L Lpinpin，ru.sru.s. .L Luiiythe.Iuiiythe.I一一.of.poi.ilseec,.of.poi.ilseec, .Sec.aysp.Sec.aysp.il.il.一一-to-toL.nuL.nurs.nLigoftensiuii,ruers.nLigoftensiuii,rue、isluinPayrrlesiteisluinPayrrlesiteL Lpispis，htse.snhtse.snr r tIebascsiiandpa-ica.qtIebascsiiandpa-ica.q -eits1.nsiuiiPayu.sliarning

13、ofthe-eits1.nsiuiiPayu.sliarningofthe”nsilli.aLLepnsilli.aLLepUILUIL. .ILILI,I,oftepaysn.ue.ulos,guiLigpioftepaysn.ue.ulos,guiLigpiple,g.ple,g.、orrgaiaton,goodsygrspcoiLiiis,htsaLoblgainsofl.ymemb.sbeaigimiorrgaiaton,goodsygrspcoiLiiis,htsaLoblgainsofl.ymemb.sbeaigimiILILt.oat,cleaLlalikLlalym-b.t.o

14、at,cleaLlalikLlalym-b. aILaILiLtonsLeaningte-opof-lLscilneIfteiLtonsLeaningte-opof-lLscilneIfte，.te.te，ssL LBCBC。a,rrgulainsteiiesmmunstPay1a,rrgulainsteiiesmmunstPay1r rb.RgsOLiaib.RgsOLiaia aILILsoon,masoon,marng-lrng-lLSLSC C|IIesanLadsof|IIesanLadsoffum-,thefuruoldigs,mlsteigfum-,thefuruoldigs,m

15、lsteigiustypsofLsiustypsofLspliayof.pliayof.a aILILpusmentprovsiisIs.ssok.ToXIpusmentprovsiisIs.ssok.ToXIier.So.ta,seis.poratsp.chier.So.ta,seis.poratsp.chdig(dig(sinandoutpov.yfrbasinandoutpov.yfrba.xtook,llariguuL.s.LXIGe.alSo.ta,s.s.por.ntsp.chofbbs.xtook,llariguuL.s.LXIGe.alSo.ta,s.s.por.ntsp.ch

16、ofbbssii,leaninguiL.sandCeetalrrlingsii,leaninguiL.sandCeetalrrlingoigpolkanewoigpolkanewn nptnewthooghtnewstptnewthooghtnewstpyofbasccnteni,.aningund.sandXIGp.ZpceayiIFu.IwrkLuingpyofbasccnteni,.aningund.sandXIGp.ZpceayiIFu.IwrkLuing 一一.of-ourgrr.of-ourgrr rots-rots-、mnayondodopal*birdfirsfy-andlhe

17、lNanpingretailsudyplposdof.gaviyXamoV,adnnov.inmlecaismmnayondodopal*birdfirsfy-andlhelNanpingretailsudyplposdof.gaviyXamoV,adnnov.inmlecaism、impolaithouggtM.impolaithouggtM.rands.ngth.tIepayspirit,ufthepurposeLoasmor.chaac.rands.ngth.tIepayspirit,ufthepurposeLoasmor.chaac.ns.ns.I Iatlnrpatdtotebasc

18、atlnrpatdtotebasc. .LUILUI. .Manund.sadm-tefloManund.sadm-teflogasecscon.nt(Lgasecscon.nt(ede,andt.ycnfidec.,andsst,andt.ycnfidec.,andsstn.nce4-fuan.nce4-fuaurSaegyurSaegyusnewofhusnewofhorycoiolions.aparyruligacigpoltc.totlStapy,cnscoosyw.orycoiolions.aparyruligacigpoltc.totlStapy,cnscoosyw.、uaul-l

19、uaul-lthework.issedinolln,aidcoordinaton,aidgee,aIdopen,adSaedoeVlpmeitsrthework.issedinolln,aidcoordinaton,aidgee,aIdopen,adSaedoeVlpmeitsrinsiplaoeVlpmetgobaoeepcaggeaccLngtnewinsiplaoeVlpmetgobaoeepcaggeaccLngtnewveopmentcic.tveopmentcic.tjob.job. pactcelneSoc-tcorevauespromo.Soci.istthoughtmoral

20、a.inpactcelneSoc-tcorevauespromo.Soci.istthoughtmorala.intaLilialvruetaLilialvrue预变性94C5min终延伸72c10min（四）PRPR产物检测：琼脂糖凝胶电泳1、电泳缓冲液TBE（10立的配制：称取Tris-base54g,硼酸27.5g,并加入0.5MEDTA（PH8.0）20mL,定容至1000mL。2、1.0%琼脂糖凝胶的配制：称取1.0g琼脂糖于三角锥形瓶中，加入10mL10kBE缓冲液，并加水定容至100mL,力口热溶解后力口入5LEB（10mg/mL）,摇匀后倒入插有梳子的电泳槽，待其凝固后拔去梳子

21、即可用于电泳。3、电泳检测：取PCR产物5NL于LoadingBuffer1pL混合， 加入到琼脂糖凝胶点样孔中，同时在样品孔旁加入5LDL2000DNAMarker作对照，以100V电压，50mA电流进行电泳，约15min后于紫外灯下观察结果。基因克隆（一）DNADNA片段与载体的连接1、反应体系（10同：组份体积LigationSolution15仙 LPCR回收产物4.5LpMD18-T0.5pL总体积102、反应条件：16C,4小时以上变性退火延伸94c55c72c1min1min1minJ：-30Cycles（二）感受态细胞的制备ulsruulsru，ueuepartpart，isa

22、ll.e.bersisall.e.berssusu”,usI,usIuarLofiatuarLofiatogetbogetb，iotseekiiotseekiureaiL.issiureaiL.issioi,furtoi,furtigguoLeigguoLe，iig,so.eeviig,so.eevlerclearfouraservilerclearfouraservieiperfuictoreiperfuictor，aLLress,evaLiaLLress,evaLiue-ofequiue-ofequie.eits,.aste.eits,.astigrespoisibilitigrespoi

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26、LLiuiliieIllLiuiliieIll，L LIe.asIe.as4ac4ac，papaiaice,suclastleiaice,suclastleiotpariiaeit.iotpariiaeit.igtleSocialistcorevaligtleSocialistcorevalOgaizaii,isOgaizaii,islotriqlielotriqlieuesFousoiseisiiuesFousoiseisiiLtopaLtopa，.e.bersli.e.bersliigeit.e.berigeit.e.berpLuesforaloigti.e,LoiotplapLuesfo

27、raloigti.e,Loiotplas,kis,kiowleowle，avaiguarLaavaiguarLai.ee.plai.ee.pla，rol,so.Lileerol,so.Lileei.eitii.eitiseeUioiai.appoi.eitofpaseeUioiai.appoi.eitofpa，a.Gover.ea.Gover.eitleaLiigcaLresworkofi.portaitleaLiigcaLresworkofi.portaigcigc，wtwtI IeiLiigpraUUeleaieiLiigpraUUeleaiigcoseigcose，weaigpracwe

28、aigpracioteioogl,ai.s.eeveioteioogl,ai.s.eeveue-i-et*Igierai.sr.toi.prue-i-et*Igierai.sr.toi.privalueivalueL-riL-riovetleiovetleii.eloi.eloois,.orois,.orlogiala.poiilogiala.poiiuU,uU,一一，v v、st,streigtleipartst,streigtleipart，spirit,listeiiigpartspirit,listeiiigpart，,part,part，,part,part，,part,part，,

29、 ,ig,focus.asterig,focus.asterCoistitutiCoistitutioi.asteraiL.e.bers,aoi.asteraiL.e.bers,apartpart，partpart，guarLiguarLi1 1g1g1ataty yaieaiep parar，1 1igtlecoisciousiessoftleigtlecoisciousiessoftleiLpartiLpart，oforgaiizatioisoforgaiizatiois，ste.,aiLpartste.,aiLpart，ofCeofCeitralOrgaitralOrgaiizaioi,

30、ai.pariizaioi,ai.par，ofpaofpaotiigrefor.,Levotiigrefor.,Levpartpart，sselop.eitaiLelop.eitaiLceorgaiizatioi,aiLpartceorgaiizatioi,aiLpart，ofgrassofgrass试剂的配制：1、LB液体培养基：称取蛋白陈（Tryptone）10g,酵母提取物（Yeastextract）5g,氯化钠10g,溶解于800mL去离子水中，用氢氧化钠调PH至7.5,加去离子水定容至1L,高温高压灭菌20min。2、0.1mol/L氯化钙溶液：称取1.11gCaC2（无水，分析纯）

31、，溶于超纯水中，定容至100mL，高温高压灭菌或0.22m滤膜过滤除菌。3、无菌甘油：取甘油100mL,高温高压即可。操作步骤（无菌操作）：1、从生长有大肠杆菌DH5a的平板上挑取一个单菌落，接种于2mLLB液体培养基中，37C200rpm,震荡培养过夜，作为一级种子。2、将一级种子按照1:50比例无菌转接到5mLLB液体培养基中，37c200rpm震荡培养约2-3小时，至细菌的OD600达至U0.5左右。3、在无菌条件下将细菌转移到一个无菌的1.5mL离心管中，冰浴30min。4、4C,4000rpm离心10min,弃上清。5、加入1mL预冷的冰的0,1mol/LCaCl2,轻轻重悬。悬浮的

32、感受态细胞可直接用于转化，若要保存，则需加入无菌甘油至终浓度为15%,分装100人/管，-70c保存备用。（三）连接产物的转化试剂配制：1、氨苇青霉素（Ampicillin,Amp）母液：用灭菌的ddH2O将氨苇青霉素粉配成100mg/mL水溶液,经0.22叩滤膜过滤除菌后，置-20C保存备用。2、Amp+抗性的LB液体培养基：在LB液体培养基中加入1人100mg/mL的氨苇青霉素母液后混匀即可， 一般现配现用。3、Amp+抗性的LB固体培养基：omaionaomaionaILILCouCCouC， 一一“ woleawoleanaLolea.araium.it.i.cl.prnaLolea.

33、araium.it.i.cl.pra.fora.forL.eu-rSaiL.gofI.Cis.uiIrU.L.eu-rSaiL.gofI.Cis.uiIrU.、HiUiIHiUiI，r-S.L0|r-S.L0| IIseesILLIIseesILLtI.baScSpiiaL|atI.baScSpiiaL|aeqeq -eitsl.C.luiI-eitsl.C.luiI，ae*laae*la，usiewusiewofIsorofIsor，coiiiioisXaparHigacigpolkaIcoiiiioisXaparHigacigpolkaIotalstrategotalstrateg，,uo

34、isui,uoisuiPayuesLiariiigofPayuesLiariiigofIIII. .pai.iipai.iili,aLeiuiLesaiLigofli,aLeiuiLesaiLigofI I. .payspaysLigpicLigpicLlob,Llob, papapr.-m.spit-orpr.-m.spit-orgig,.gig,._ _g.,rs,eiueeariiga-uaioi.g.,rs,eiueeariiga-uaioi.“eecve.sspro.oeal.eecve.sspro.oeal.bebescisiousyscisiousy*gI*gILsigusleL

35、siguslea aiL11stageiL11stageelieSoc-tcorevauespro.oeSocilstIlooglt.oraelieSoc-tcorevauespro.oeSocilstIlooglt.oralaLCiieela.iiialviu.laLCiieela.iiialviu.ousous，es.UILCisiuloi,es.UILCisiuloi,IparIpar，rus,wilxirus,wilxi，eaea、uetaueta，seesirnportaitspecsiia.Lseesirnportaitspecsiia.L.L L, ,guaiisI.Ciiee.

36、usPalguaiisI.Ciiee.usPal，1 1r rbeRigsOrLiaiceaiLsooi,mabeRigsOrLiaiceaiLsooi,ma.igielLsUpinsa.a.igielLsUpinsa.a.igs,.igs,.IeiIeie.eitsaree.eitsareleelee，.aksIefol.aksIefolowigspeUfcproga.owigspeUfcproga.- -LeaiiLeaiiigreui-esoiaigreui-esoialparlpar， . .besIbesIostuLostuL，tIeparttIepart，Coistitutioia

37、iLCoistitutioiaiLpartpart，rulerulesaLuii.iieXi-resof.poraitseesaLuii.iieXi-resof.poraitseeu,u, eaeaS Sueauea，spespeUlariig-r-IoUlariig-r-IosUpliasUplia ， ofeieaofeieaILILpuis.eitprovsoisIseiessokeToXiXieaSiuelapuis.eitprovsoisIseiessokeToXiXieaSiuela ， sese.poratspeclradig.poratspeclradig （ esiiB.-l

38、es.-clyB.I Ia aLOLO_ _I I_ _11garai.me.speclkpr11garai.me.speclkpr.me.me.-ul.-ul”yrUeparyLlgtyrUeparyLlgt，.Ogaibaton.entonCuy,.Ogaibaton.entonCuy,I Ia aLOLO_ _I I_ _11garai.me.ofseclkpro11garai.me.ofseclkpro”.,.,. .L LIBoaIBoaL L-rib.L-Hi.p.m.itiaonp.-rib.L-Hi.p.m.itiaonp.espit.espitordigt.ordigt.”.

39、 .a aILILh-th-t”.Ulip.Ulip.of.i-.ts,.nu.a.ing.of.i-.ts,.nu.a.ing.ion.ion. .*.*promo.alm-b.scisl.*.*promo.alm-b.scisl.esectdi.esectdi.“on,aion,aiL L-.y-.y- -aruaru,wt.iGe.c.a,wt.iGe.c.a，se.s.por.ntse.s.por.nt一一silsil.I.IL L,.,.L L.pracbeaipracbeaiL LPromo.teworkaPromo.teworkaILILbequOi-pa.bequOi-pa.b

40、e.be. .L L.ilg.ng.ilg.ng.eh.eh.，mamatefolo-Wigspeciepr.mm-.L-.ing.-I-.tefolo-Wigspeciepr.mm-.L-.ing.-I-.、01alpa.01alpa.be.t.be.t.，.payi.In.payi.In. .L Lpinpin，ru.sru.s. .L Luiiythe.Iuiiythe.I 一一.of.poi.ilseec,.of.poi.ilseec, .Sec.aysp.c.Sec.aysp.cilg-ilg-to-toL.nuL.nurs.nLigoftensiuinruers.nLigoften

41、siuinrue、nsiuinP.yrrlesitensiuinP.yrrlesiteL L- -pispis，htse.sidiLlestebascsiiandp.-ica.qhtse.sidiLlestebascsiiandp.-ica.q -eits1.nsiuinP.yu.sLLarningofthe-eits1.nsiuinP.yu.sLLarningofthe”nsilli.aLL.pnsilli.aLL.pUILUIL. .ILILI,I,oftepaysn.ueoftepaysn.ue ulos,guiLigpiulos,guiLigpip.,g.p.,g.、orrgaiato

42、n,goodsy.grspconiHins,hts.Loblgainsofl.ymemb.sbe.igimiorrgaiaton,goodsy.grspconiHins,hts.Loblgainsofl.ymemb.sbe.igimiILILt.oat,cleat.oat,cleaQ QIBIBL L,.,m,.,m b.b.一一aILaILILIL! !onsLeaningte-L.of-lLscilneIfteonsLeaningte-L.of-lLscilneIfte，.te.te，sLisClk.,rrgulainsteinne.mmunstP.y1sLisClk.,rrgulains

43、teinne.mmunstP.y1r r-b.RgsOLiai-b.RgsOLiaia aILILsoon,masoon,marng-lLisC|IIesaiL.rsoffumus,thefuruholdigs,mlsteigrng-lLisC|IIesaiL.rsoffumus,thefuruholdigs,mlsteigiustypsofLsiustypsofLspliayof.neapliayof.neaILILpuismentprovsiisIs.ssok.ToXIXner.So.ta,s.is.por.tsp.chpuismentprovsiisIs.ssok.ToXIXner.So

44、.ta,s.is.por.tsp.chdigdig（sinandoutpov.yfrbasinandoutpov.yfrba.xtook,llariguuL.s.LXIGe.alSo.ta,s.s.por.ntsp.chofbbs.xtook,llariguuL.s.LXIGe.alSo.ta,s.s.por.ntsp.chofbbssii,.ningunL.sasii,.ningunL.saILILCeet.lrrlngCeet.lrrlngoigpolk.new-in.ptnewthooghtnewsta.gyofbasoigpolk.new-in.ptnewthooghtnewsta.g

45、yofbasnteni,.aningund.mndXIG.2.-e.yiIFu.nteni,.aningund.mndXIG.2.-e.yiIFu.n nwrkLuingwrkLuing（dvocaeofourgrasrottmmidiaeyondodopadbirdfirsfyandlhelNanpingreeaclsudyplposdofgaliyXamoV,adnnov.ion.CaismdvocaeofourgrasrottmmidiaeyondodopadbirdfirsfyandlhelNanpingreeaclsudyplposdofgaliyXamoV,adnnov.ion.C

46、aism、impol.Ithouggt*impol.Ithouggt*rands.ngth.tIep.yspirit,ulflsthepurposeLiasmor.chaa-.rands.ngth.tIep.yspirit,ulflsthepurposeLiasmor.chaa-.isevtonrisevtonrtotebastotebas.qur.qur.tlManuid.s.dmistefloWngase-scon.nt.tlManuid.s.dmistefloWngase-scon.nt（ ,andpeopleofd.0,isinemmodoylamosgratofdrere,andpe

47、opleofd.0,isinemmodoylamosgratofdre,-reEs.ntasisI.ion.pros.,-reEs.ntasisI.ion.pros.、adn.ionalreVtalzaton,adpeopadn.ionalreVtalzaton,adpeop，eh.piineseh.piines；baffe.u.sSo-.isisabaffe.u.sSo-.isisabeedinnseg.beedinnseg.lalIfway.hancedroad-nld.lalIfway.hancedroad-nld.,andt.y-nfidec.,andsst,andt.y-nfidec

48、.,andsstnld.nld.e4fuae4fuaurSaegyurSaegyusnewofhisor,coniltionsX.parulngactngpoltcatotlst.ey,-ns-oosywitfuaulusnewofhisor,coniltionsX.parulngactngpoltcatotlst.ey,-ns-oosywitfuaulthework.bssedlnoaton,andcoordinaton,andgee,aIdopen,adsa.dthework.bssedlnoi.,wt.iGe.i.，seesbmpor.ntspei.seesbmpor.ntspei.IL

49、ILmb.,mb.,. .L L”prapraaiaiL LPro.oeteworkaPro.oeteworkaILILbequOibequOipa.pa.be.be. .L Le.ibg.nge.ibg.ng.eh.eh.，mamatefolo-Wigspeclipr.mm-.I_ilngr-b-e01alpa.tefolo-Wigspeclipr.mm-.I_ilngr-b-e01alpa.be.t.be.t.，.pay.pay，.i.i.“i i. .L Lpinpin，.s.s. .L LLilythe.ILilythe.I.ofbmpoi.nseeI,.ofbmpoi.nseeI,

50、.Sei.ayspel.Sei.ayspelibg-ibg-to-toL.nuL.nurs.iLbgofthensbutonruesC.istlubnPalyrrlesbteLL.pbsrs.iLbgofthensbutonruesC.istlubnPalyrrlesbteLL.pbs， htse.snhtse.snr r tIebas.spbbaLpa111a.Llbroeits1.nsbutbiPalyu.sliarnbigofthetIebas.spbbaLpa111a.Llbroeits1.nsbutbiPalyu.sliarnbigofthe”nstllloi.aLL.pLlL.Sa

51、lLilgoftepaysn.ue.ulos,gubLbigpblp.sgg.sorga-ton,goodsypnstllloi.aLL.pLlL.SalLilgoftepaysn.ue.ulos,gubLbigpblp.sgg.sorga-ton,goodsypG GP Ploiibbiloiibbi， ， hts.Loblhts.Lobl， abisof,aymemb.sbeabgbnmbndt.oat,ileaLlalHeLlalymob.abisof,aymemb.sbeabgbnmbndt.oat,ileaLlalHeLlalymob. aILuiLbtonsLaILuiLbtons

52、Llilgtheioeof-lUsUplneIfthelilgtheioeof-lUsUplneIfthe， ， ， .te.te， ， ，sLbsllbayrrgulabisthesLbsllbayrrgulabistheIIIIIBIIIBCommubsPaly1CommubsPaly1r r-b.RghtsOrbnaineandsoon,ma-b.RghtsOrbnaineandsoon,marng-lnsl|IIerng-lnsl|IIeILIL.-.-offum-,thefuruholdbgs,m-tebgoffum-,thefuruholdbgs,m-tebgbustypsofLb

53、slplbayof.neandpubsmentprovsonsIs.bessok.ToXIi.aSn.ta,s.mbmporatsp.ihbustypsofLbslplbayof.neandpubsmentprovsonsIs.bessok.ToXIi.aSn.ta,s.mbmporatsp.ihLbgLbg（-sbn-sbn）aiLoutpov.yfrbasiaiLoutpov.yfrbasitook,llanbiguners.LXIGe.alStook,llanbiguners.LXIGe.alS.ta,s.besbmpor.itsp.ihobslsH,leanbiguiL.sandCee

54、talrrlng.ta,s.besbmpor.itsp.ihobslsH,leanbiguiL.sandCeetalrrlngolbgpolbcanewinn.ptnewthooghtnewstrtepyofbasiinteni,.aibigund.sandXIGp.,irtaybIFu.Iwrkdubigmvoiaeof_ourgrrolbgpolbcanewinn.ptnewthooghtnewstrtepyofbasiinteni,.aibigund.sandXIGp.,irtaybIFu.Iwrkdubigmvoiaeof_ourgrr .otsmmmbaeyondonopa.otsm

55、mmbaeyondonopa，bbrdfbrsfy_aidlhebiNaipbigre.ausudyp.poslof.galbyXamold,aidnnov.bonmeUabsmbbrdfbrsfy_aidlhebiNaipbigre.ausudyp.poslof.galbyXamold,aidnnov.bonmeUabsm、bmpod.Ithouggt*seraids.ngth.tIepayspbrbt,uflsthepurposebdrasmor.ihaaierinsbmpod.Ithouggt*seraids.ngth.tIepayspbrbt,uflsthepurposebdrasmo

56、r.ihaaierinsabonreattotebasi.Lurabonreattotebasi.Lur.tsUnund.sadmlsteflo.tsUnund.sadmlsteflogaseision.ntgaseision.nt（bdelfbthbsmmunsss,Shangofalbum,s.rbghtofwold.ewadOulookonlf.aidB.-les.-clyB.I Ia aLOLO_ _I I_ _11garai.me.speclkpr11garai.me.speclkpr.me.me.-ul.-ul”yrUeparyLlgtyrUeparyLlgt，.Ogaibaton

57、.entonCuy,.Ogaibaton.entonCuy,I Ia aLOLO_ _I I_ _11garai.me.ofseclkpro11garai.me.ofseclkpro”.,.,. .L LIBoaIBoaL L-rib.L-Hi.p.m.itiaonp.-rib.L-Hi.p.m.itiaonp.espit.espitordigt.ordigt.”. .a aILILh-th-t”.Ulip.Ulip.of.i-.ts,.nu.a.ing.of.i-.ts,.nu.a.ing.ion.ion. .*.*promo.alm-b.scisl.*.*promo.alm-b.scisl

58、.esectdi.esectdi.“on,aion,aiL L-.y-.y- -aruaru,wt.iGe.c.a,wt.iGe.c.a，se.s.por.ntse.s.por.nt一一silsil.I.IL L,.,.L L.pracbeaipracbeaiL LPromo.teworkaPromo.teworkaILILbequOi-pa.bequOi-pa.be.be. .L L.ilg.ng.ilg.ng.eh.eh.，mamatefolo-Wigspeciepr.mm-.L-.ing.-I-.tefolo-Wigspeciepr.mm-.L-.ing.-I-.、01alpa.01al

59、pa.be.t.be.t.，.payi.In.payi.In. .L Lpinpin，ru.sru.s. .L Luiiythe.Iuiiythe.I 一一.of.poi.ilseec,.of.poi.ilseec, .Sec.aysp.c.Sec.aysp.cilg-ilg-to-toL.nuL.nurs.nLigoftensiuinruers.nLigoftensiuinrue 、 nsiuinP.yrrlesitensiuinP.yrrlesiteL L- -pispis ， htse.sidiLlestebascsiiandp.-ica.qhtse.sidiLlestebascsiia

60、ndp.-ica.q -eits1.nsiuinP.yu.sLLarningofthe-eits1.nsiuinP.yu.sLLarningofthe”nsilli.aLL.pnsilli.aLL.pUILUIL. .ILILI,I,oftepaysn.ueoftepaysn.ue ulos,guiLigpiulos,guiLigpip.,g.p.,g. 、 orrgaiaton,goodsy.grspconiHins,hts.Loblgainsofl.ymemb.sbe.igimiorrgaiaton,goodsy.grspconiHins,hts.Loblgainsofl.ymemb.sb

61、e.igimiILILt.oat,cleat.oat,cleaQ QIBIBL L,.,m,.,m b.b. 一一 aILaILILIL! !onsLeaningte-L.of-lLscilneIfteonsLeaningte-L.of-lLscilneIfte ， ， ， ， ， .te.te ， ， ， ， ，sLisClk.,rrgulainsteinne.mmunstP.y1sLisClk.,rrgulainsteinne.mmunstP.y1r r-b.RgsOLiai-b.RgsOLiaia aILILsoon,masoon,marng-lLisC|IIesaiL.rsoffumu

62、s,thefuruholdigs,mlsteigrng-lLisC|IIesaiL.rsoffumus,thefuruholdigs,mlsteigiustypsofLsiustypsofLspliayof.neapliayof.neaILILpuismentprovsiisIs.ssok.ToXIXner.So.ta,s.is.por.tsp.chpuismentprovsiisIs.ssok.ToXIXner.So.ta,s.is.por.tsp.chdigdig（sinandoutpov.yfrbasinandoutpov.yfrba.xtook,llariguuL.s.LXIGe.al

63、So.ta,s.s.por.ntsp.chofbbs.xtook,llariguuL.s.LXIGe.alSo.ta,s.s.por.ntsp.chofbbssii,.ningunL.sasii,.ningunL.saILILCeet.lrrlngCeet.lrrlngoigpolk.new-in.ptnewthooghtnewsta.gyofbasoigpolk.new-in.ptnewthooghtnewsta.gyofbasnteni,.aningund.mndXIG.2.-e.yiIFu.nteni,.aningund.mndXIG.2.-e.yiIFu.n nwrkLuingwrkL

64、uing（dvocaeofourgrasrottmmidiaeyondodopadbirdfirsfyandlhelNanpingreeaclsudyplposdofgaliyXamoV,adnnov.ion.CaismdvocaeofourgrasrottmmidiaeyondodopadbirdfirsfyandlhelNanpingreeaclsudyplposdofgaliyXamoV,adnnov.ion.Caism、impol.Ithouggt*impol.Ithouggt*rands.ngth.tIep.yspirit,ulflsthepurposeLiasmor.chaa-.r

65、ands.ngth.tIep.yspirit,ulflsthepurposeLiasmor.chaa-.isevtonrisevtonrtotebastotebas.qur.qur.tlManuid.s.dmistefloWngase-scon.nt.tlManuid.s.dmistefloWngase-scon.nt（ ,andpeopleofd.0,isinemmodoylamosgratofdrere,andpeopleofd.0,isinemmodoylamosgratofdre,-reEs.ntasisI.ion.pros.,-reEs.ntasisI.ion.pros.、adn.i

66、onalreVtalzaton,adpeopadn.ionalreVtalzaton,adpeop，eh.piineseh.piines；baffe.u.sSo-.isisabaffe.u.sSo-.isisabeedinnseg.beedinnseg.lalIfway.hancedroad-nld.lalIfway.hancedroad-nld.,andt.y-nfidec.,andsst,andt.y-nfidec.,andsstnld.nld.e4fuae4fuaurSaegyurSaegyusnewofhisor,coniltionsX.parulngactngpoltcatotlst.ey,-ns-oosywitfuaulusnewofhisor,coniltionsX.parulngactngpoltcatotlst.ey,-ns-oosywitfuaulthework.bssedlnoaton,andcoordinaton,andgee,aIdopen,adsa.dthework.bssedlnoi.,wt.iGe.i.，seesbmpor.ntspei.seesbmpo