简介:Themodulationandcontrolofgecko'sfootmovementswerestudiedelectrophysiologicallyinordertodesignthemotorcontrolsystemofagecko-mimicrobot.Inthisstudy(1)theanatomyoftheperipheralnervescontrollingthegecko'sfootmovementswasdetermined;(2)therelationshipbetweenthelimbnervesofthegeckoanditsfootmotorpatternswasstudied;(3)theafferentimpulsesofthenervesevokedbyrubbingthegecko'stoesandpalmwererecorded;(4)copyingthenaturalpatternsofmovementofthegecko'sfoot(abduction,adduction,flexion,andrevolution)anditslimbnervemodulationandcontrolmechanism,thenerveswerestimulatedundercomputercontrol,andtheresultsrecordedbyCCD.Resultssuggestthatgecko'sfootmovementscanbesuccessfullycontrolledbyartificialelectricalsignals.
简介:Thesandfishisalizardhavingtheremarkableabilitytomoveindesertsandinaswimming-likefashion.Themostout-standingadaptationstothismodeoflifearethelowfrictionbehaviourandtheextensiveabrasionresistanceofthesandfishskinagainstsand,outperformingevensteel.Weinvestigatedthetopography,thecompositionandthemechanicalpropertiesofsandfishscales.Theseconsistofglycosylatedkeratinswithhighamountofsulphurbutnohardinorganicmaterial,suchassilicatesorlime.Remarkably,atomicforcemicroscopyshowsanalmostcompleteabsenceofattractiveforcesbetweenthescalesurfaceandasilicontip,suggestingthatthisisresponsiblefortheunusualtribologicalproperties.Theunusualglycosylationofthekeratinswasfoundtobeabsolutelynecessaryforthedescribedphenomenon.Thescalesweredissolvedandreconstitutedonapolymersurfaceresultinginpropertiessimilartotheoriginalscale.Thus,weprovideapathwaytowardsexploitationofthereconstitutedscalematerialforfutureengineeringapplications.
简介:Naturalcomposites,formedthroughbiomineralization,havehighlyorderedstructureswhichhavebeenaptlyexploredforfunctionalapplications.Thoughtheroleoforganicphaseshasbeenwellunderstoodinbiomineralization,notenoughattentionhasbeenpaidtotheroleofbio-membraneswhichareoftenfoundencapsulatingthechamberinwhichmineralizationoccurs.Wehaveusedthenaturalproteinandsemi-permeablemembraneofchickeneggstogrowdifferentmaterialssuchasceramics,semi-metalsandmetalstounderstandtheroleofbio-membranesinbiomineralization.Weherereportthesuccessfulbiomimeticsynthesisofcalcite,cadmiumsulphide,andsilverhavinghomogeneousmorphologies.Wehavefoundthatthemembraneoperateslikeatunedgateway,playingasignificantroleincontrollingthemorphologyoftheinorganiccrystalsformedduringbiomineralization.
简介:目的:应用重组杆状病毒表达系统制备由HA、NA、M1和M2蛋白组成的H5N1高致病性禽流感病毒样颗粒,为研究H5N1高致病性禽流感疫苗奠定基础。方法:构建能共表达A/chicken/Jilin/2003(H5N1)禽流感病毒血凝素(HA)和神经氨酸酶(NA)、A/PR/8/34(H1N1)流感病毒基质蛋白(M1)和离子通道蛋白(M2)的2个二元重组杆状病毒,共同感染HighFive细胞,同时表达HA、NA、M1和M2蛋白,使这4种蛋白在感染的细胞内自主组装成病毒样颗粒。经差速离心和蔗糖密度梯度超速离心收获病毒样颗粒,通过Western印迹鉴定病毒样颗粒的组成,透射电镜观察病毒样颗粒形态,血凝试验测定病毒样颗粒的活性。结果:HA、NA、M1、M2蛋白在昆虫细胞中共表达,并组装成病毒样颗粒;电镜观察到病毒样颗粒的形态与流感病毒一致,直径约80nm;血凝试验显示该病毒样颗粒具有凝集鸡红细胞的活性。结论:应用该方法可以制备流感病毒样颗粒,为H5N1流感疫苗研究提供了可行方案。
简介:目的:构建40S核糖体蛋白S6的原核表达载体,表达并纯化S6蛋白,将其作为底物用于S6激酶(S6K)的体外活性测定。方法:采用RT-PCR方法从人胚肾细胞HEK293中获取S6cDNA,将扩增产物克隆至大肠杆菌表达载体中,进行酶切及测序鉴定;IPTG诱导GST-S6融合蛋白在大肠杆菌中表达,用谷胱甘肽亲和层析纯化GST-S6,免疫沉淀法检测该蛋白是否可作为底物用于S6K的体外激酶活性测定。结果:酶切及测序鉴定表明构建了S6原核表达载体,并表达及纯化出GST-S6融合蛋白,相对分子质量为55×103。该蛋白可用于S6K的体外激酶活性测定,特异性强。结论:S6蛋白的克隆、表达与纯化成功,可用于S6K的体外激酶活性测定,为研究S6K的功能奠定了基础。
简介:目的:分离羽衣甘蓝S13-b位点受体激酶(SRK13-b)基因并进行序列及结构域分析,构建SRK13-b结构域的原核表达载体并进行重组蛋白质的原核表达和纯化。方法:提取羽衣甘蓝S13-bS13-b自交不亲和系花期柱头的RNA,用RT-PCR法分离SRK13-b基因;将编码SRK13-b激酶结构域的序列插入大肠杆菌表达载体pET-14b中,构建原核表达质粒pET-SRK13-bCT,转化大肠杆菌BL21(DE3)pLysS菌株,经0.1mmol/LIPTG诱导,用Ni—NTA亲和层析柱对SRK13-b激酶结构域蛋白进行纯化。结果:分离获得羽衣甘蓝SRK13-b基因的长度为2571bp,编码856个氨基酸,GenBank收录号为EU180597;对SRK13-b激酶结构域蛋白进行诱导表达及纯化,SDS—PAGE显示相对分子质量约43×10^3的蛋白质特异表达,对表达产物进行分离纯化,获得了SRK13-b激酶结构域的融合蛋白。结论:羽衣甘蓝SRK13-b基因的克隆及激酶结构域的原核表达,为研究SRK的功能及自交不亲和性奠定了基础。
简介:目的:构建一个牛dSl酪蛋白调控序列指导人溶菌酶(hLYZ)基因组序列的杂合基因座。方法:采用本实验室发明的连续3步缺口修复技术。将6个无痕连接的同源臂插入以pBR322为载体的骨架中,构成能进行3次连续基因抓捕的载体,利用Red同源重组系统介导的缺口修复技术,分别抓捕牛aSl酪蛋白3’端调控序列(9kb)、hLYZ基因座序列(5kb)、牛axSl酪蛋白5’端调控序列(20kb),使这3个基因片段自动无痕地连接在基因抓捕载体上,形成牛oLSl酪蛋白一hLYZ杂合基因座。结果:实验经过PCR扩增、限制性内切酶酶切验证和序列测定,验证了hLYZ的基因组序列对牛仪S1酪蛋白编码基因组序列的精确置换。结论:这种修复技术为乳腺生物反应器高效表达大载体的制备提供了可行的思路及方法。