Cloning of TPA Gene and Establishing of Expressing Model in vitro / 中国医师杂志

ABSTRACT

Objective To recombine the tissue plasminogen activator (TPA) gene and establish the expressing model in vitro to provide both theoretical basic and technical guidance for gene therapy of ischemic heart disease and prevention of postoperative vessel re-stenosis. Methods Expression vector pcDNA3.1 TPA was constructed, and transfected into Chinese hamster ovary(CHO) cells. Then the exogenous TPA expression was observed. Results The expression quantity of TPA in transfected CHO cells was higher than that in non-transfected cells. The exogenous TPA activity was 12 296 IU/10 6cell/24hr in the transfected cells, when measured by chromogenic substrate assay, while that of non-transfected cells was 3 176IU/10 6cell/24hr. The TPA quantity of non-transfected cells was 9 608 ng/10 6cell/24hr, when measured by enzyme-linked immunosorbent assay (ELISA), while that of transfected cells was 586 172 ng/10 6 cell/24hr, and the latter was the 60 times of the former. Conclusions When pcDNA3 1(+)TPA was transfected into CHO cells, exogenous TPA can efficiently be expressed, which provides theoretical basic for TPA clinical gene therapy.