|
OALib Journal期刊
ISSN: 2333-9721
费用：99美元
|
|
|
|
人可溶性晚期糖基化终产物受体的原核表达及活性鉴定
, PP. 769-773
Keywords: 晚期糖基化终产物,原核表达,受体,受体,可溶性,融合蛋白/生物活性
Abstract:
目的构建人可溶性晚期糖基化终产物受体(hsrage)his融合蛋白表达载体并在原核细胞表达与纯化。方法pcr扩增hrage基因编码区的胞质外段,利用分子克隆技术将其重组于pet-14b载体中。利用酶切、序列分析鉴定克隆正确性。转化大肠杆菌bl21(de)3,用异丙基b-d硫代半乳糖(iptg)诱导融合蛋白表达。以尿素对获得的包涵体进行处理,用金属螯合亲和层析的方法纯化融合蛋白并进行复性。以western印迹和elisa法对融合蛋白的免疫原性及与age的结合活性进行鉴定。结果克隆的hsrage完全正确,经过表达与纯化,获得了相对分子质量约45000的融合蛋白。纯化得到的变性蛋白经复性后可被相应抗体识别。所得到的hsrage具有与age相互结合的活性,并能够抑制age诱导的内皮细胞nf-kb的表达。结论成功地构建了hsrage融合蛋白原核表达载体且获得高效表达,得到大量的复性蛋白;该蛋白具有特异的免疫原性,保持与age的结合活性,并能够有效阻断age-rage相互作用。
| [1] | houff,jiangjp,guojq,etal.receptorforadvancedglycationendproductsonhumansynovialfibroblasts:roleinthepathogenesisofdialysis-relatedamyloidosis[j].jamsocnephrol,2002,13:1296-130
|
| [2] | katzj,bhattacharyyai,farkhondeh-kishf,etal.expressionofthereceptorofadvancedglycationendproductsingingivaltissuesoftype2diabetespatientswithchronicperiodontaldisease:astudyutilizingimmunohistochemistryandrt-pcr[j].jclinperiodontol,2005,32(1):40-4.
|
| [3] | 赵善超,刘靖华,李志杰,等.人晚期糖基化终产物受体胞质内段融合蛋白表达载体的构建与表达[j].第一军医大学学报,2003,23(12):1314-6.zhaosc,liujh,lizj,etal.vectorconstructionfortheintracellulardomainofhumanreceptorforadvancedglycationend-productsandexpressionofthefusionprotein[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2003,23(12):1314-6.
|
| [4] | houff,chertowgm,kayj,etal.interactionbetweenbeta2-microglobulinandadvancedglycationendproductsinthedevelopmentofdialysisrelated-amyloidosis[j].kidneyint,1997,51(5):1514-9
|
| [5] | 鄂征.组织培养和分子细胞学技术[m].北京出版社,1999.125-6.
|
| [6] | wear-maggittik,leej,conejeroa,etal.useoftopicalsrageindiabeticwoundsincreasesneovascularizationandgranulationtissueformation[j].annplastsurg,2004,52(5):519-21.
|
| [7] | bartlingb,hofmannhs,weigleb,etal.down-regulationofthereceptorforadvancedglycationend-products(rage)supportsnon-smallcelllungcarcinoma[j].carcinogenesis,2005,26(2):293-301.
|
| [8] | neeperm,schmidtam,brettj,etal.cloningandexpressionofacellsurfacereceptorforadvancedglycosylationendproductsofprotein[j].jbiolchem,1992,267(21):14998-5004.
|
| [9] | 赵善超,侯凡凡,姜勇.晚期糖基化终产物受体的结构和功能[j].生理科学进展,2003,34(1):71-3zhaosc,houff,jiangy.thestructureandfunctionofrage[j].progphysiolsci,2003,34(1):71-3.
|
| [10] | 朱平,唐磊,赵善超,等.抗人晚期糖基化终产物受体胞外段不同表位单克隆抗体的制备[j].第一军医大学学报,2004,24(2):129-32.zhup,tangl,zhaosc,etal.preparationofmonoclonalantibodiesagainstdifferentepitopesonextracellulardomainofhumanreceptorforadvancedglycationendproduct[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2004,24(2):129-32.
|
| [11] | 龚小卫,秦清河,王静珍,等.p38mapk红色荧光蛋白融合载体的构建及表达[j].第一军医大学学报,2002,22(2):171-3gongxw,qinqh,wangjz,etal.constructionandexpressionofredfluorescentproteinfusionvectorincorporatingp38mitogen-activatedproteinkinase[j].jfirstmilmeduniv/diyijunyidaxuexuebao,2002,22(2):171-3.
|
| [12] | ernstb,fortunatos,dietmars.influenceofreducedintracellularglutathioneavailabilityonthesecretionofvasoactivesubstancesbyhumanumbilicalveinendothelialcells[j].hypertenspregnancy,2001,20(1):45-56.
|
| [13] | chavakist,bierhausa,nawrothpp,etal.rage(receptorforadvancedglycationendproducts):acentralplayerintheinflammatoryresponse[j].microbesinfect.2004,6(13):1219-25.
|
| [14] | bastag,lazzerinig,massarom,etal.advancedglycationendproductsactivateendotheliumthroughsignal-transductionreceptorrage:amechanismforamplificationofinflammatoryresponses[j].circulation,2002,105(7):816-22.
|
| [15] | bucciarellilg,wendtt,quw,etal.rageblockadestabilizesestablishedatherosclerosisindiabeticapolipoproteine-nullmice[j].circulation,2002,106(22):2827-35.
|
| [16] | zengs,feirtn,goldsteinm,etal.blockadeofreceptorforadvancedglycationendproduct(rage)attenuatesischemiaandreperfusioninjurytotheliverinmice[j].hepatology,2004,39(2):422-32.
|
Full-Text
|
|
Contact Us
service@oalib.com QQ:3279437679 
WhatsApp +8615387084133
|
|
