将含完整阅读框架的人t PA (tissue typeplasminogenactivator,t PA)cDNA克隆至昆虫细胞表达转移载体pBacPAK8中 ,获得重组质粒pBac tPA .应用脂质体共转染法 ,将pBac tPA和线性化杆状病毒BacPAK6DNA共转染Tn 5B 1昆虫细胞 .经空斑筛选获得 11株重组病毒 .经PCR鉴定与生物活性测定 ,获得一株高效表达t PA的重组病毒BactPA3.t PA表达活性在感染 (MOI≈ 10 )后 72h左右达到最高 ,为 3 0 4× 10 3IU/mL .经SDS PAGE纤维蛋白自显影分析 ,相对分子质量为 680 0 0左右 . The t PA (t PA) cDNA with complete reading frame was cloned into the insect cell expression vector pBacPAK8 to obtain the recombinant plasmid pBac tPA.Using liposome co-transfection method, pBac tPA and linearized rod- The virus BacPAK6DNA was co-transfected into Tn 5B 1 insect cells, and 11 recombinant viruses were screened by plaque screening.A recombinant adenovirus with high expression of t PA was obtained by PCR identification and bioassay.t PA expression activity in infected (MOI≈ 10) reached the highest level at about 72h, which was 340 × 10 3 IU / mL.After SDS PAGE fibrinogen imaging analysis, the relative molecular mass was about 680 0 0.