You Position: Home > Paper
Construction of Expression Vector pcDNA3.1(+)TPA and Study of Its Expression Activity in Human Umbilical Vein Endotheliocytes
( views:36, downloads:0 )
- Author:
- No author available
- Journal Title:
- JOURNAL OF CHINESE PHYSICIAN
- Issue:
- 7
- DOI:
- 10.3760/cma.j.issn.1008-1372.2004.07.004
- Key Word:
- 组织型纤溶酶原激活因子;基因治疗;pcDNA3.1;脐静脉内皮细胞;缺血性心脏病
Abstract： 目的建立TPA基因在人脐静脉内皮细胞外源性表达的方法,为缺血性心脏疾病的基因治疗及防止血管再狭窄提供理论依据和新方法.方法构建真核表达载体pcDNA3.1(+)TPA,并采用脂质体法将其转入体外培养的人脐静脉内皮细胞,并观察外源性TPA表达情况.结果真核表达载体pcDNA3.1(+)TPA在人脐静脉内皮细胞中获有效表达,酶联免疫吸附法TPA蛋白表达定量检测结果为568.6ng/106细胞/24h,未转pcDNA3.1(+)TPA的人脐静脉内皮细胞测得为17.8ng/106细胞/24h;发色底物法测得外源性TPA活性为108.8IU/106细胞/24h,未转pcDNA3.1(+)TPA的人脐静脉内皮细胞测得为5.6IU/106细胞/24h.结论 pcDNA3.1(+)TPA转入人脐静脉内皮细胞后,外源性TPA基因获有效表达,为缺血性心脏病的TPA基因治疗提供了理论依据和新方法.
- [1]Saxena R,Shenaq SM,Thung S,etc.Gene therapy to promote thromboresistance: local overexpression of tissue plasminogen activator to prevent arterial thrombosis in an in vivo rabbit model.[J].1999,96(3).
- [2]C, Rosnoblet,U M, Vischer,R D, Gerard,etc.Storage of tissue-type plasminogen activator in Weibel-Palade bodies of human endothelial cells.[J].1999,19(7).
- [3]C M, Gibson,C P, Cannon,S A, Murphy,etc.Weight-adjusted dosing of TNK-tissue plasminogen activator and its relation to angiographic outcomes in the thrombolysis in myocardial infarction 10B trial. TIMI 10B Investigators.[J].1999,84(9).