RGD-Ins蛋白结构、活性和其基因治疗血栓研究.pdf

：’I．。、一忆睑义 捕婴 血糖值和血清中胰岛素样物质的含量等指标的变化，实验结果表明转入 RGD—Cys—Ins基因后，小鼠的出血时间明显延长，而血清中胰岛素样物质含量及血 糖值与转空载体的对照组比较则没有改变。同时我们用RT-PCR的方法检测到在转 基因有可能被开发成一种新的治疗和预防血管栓塞的药物。当然，要想使 RGD—Cys—Ins基因最终成为一种预防血栓的治疗基因应用于临床，还需要做进一步 工作来积累数据。 关键词：出血时间，二硫键，抑制血小板聚集，胰岛素原骨架，肽的展示 RGD功能序列，基因治疗 Abstracl Abstract Protein were to theknown engineeringtechniquesemployedgraft the onto surface anticoagulantArg—Gly-Asp(RGD)motif—containingsequences ofamutant，inactiveinsulinframework．Totheeffectofdisulfide probe bondOntheresultant native anticoagulantactivity，aRGD—containing fromavenom wellasa sequence protein modified eachexamined．Thewas sequence，SFT删PGPYS，werepeptideplaced betweentheC‘。terminaloftheBchainandtheN——terminaloftheAchainand connectedwithB27andA1 of residuestheinactiveinsulinthatlacksthe characteristicintramolecularA6—11 bondwithinA disulfide thechain．The two insulin were inE RGD—containinggenes over—expressedcull,and and as and purifled designatedRGD-Cys—Ins aminoacid andmassdatawerein with of those compositions goodagreement showed with expected．The inhibitionof RGD—Cys—Ins plateletaggregation an of thelatter IC50 3州，while was3．5