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Cloning of TPA Gene and Establishing of Expressing Model in vitro
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- Author:
- No author available
- Journal Title:
- JOURNAL OF CHINESE PHYSICIAN
- Issue:
- 11
- DOI:
- 10.3760/cma.j.issn.1008-1372.2003.11.003
- Key Word:
- 组织型纤溶酶原激活因子;CHO细胞;缺血性心脏病
Abstract： 目的重组TPA基因并构建体外表达模型,为缺血性心脏疾病的基因治疗及防止血管再狭窄奠定基础.方法构建真核表达载体pcDNA3.1(+)TPA,然后将pcDNA3.1(+)TPA转入中国苍鼠卵巢(CHO)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CHO细胞表达量好,发色底物法测得外源性TPA活性为12.296 IU/106细胞/24hr,未转pcDNA3.1(+)TPA的CHO细胞测得为3.176 IU/106细胞/24hr;酶联免疫实验(ELASA)检测结果为586.172 ng/106细胞/24hr,未转pcDNA3.1(+)TPA的CHO细胞测得为9.608 ng/106细胞/24hr,达未转TPA基因组的60倍.结论 pcDNA3.1(+)TPA转入CHO细胞后,外源性TPA基因获有效表达,为TPA临床基因治疗提供了理论依据.
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