将增强的绿色荧光蛋白 ( enhanced green fluorescent protein,EGFP)基因插在 HCMV( hum ancytom egolovirus)启动子下游 ,构建了表达质粒 p CA13 - e G,用脂质体 L ipofectin介导分别转染 He L a细胞、Vero细胞 ,仅通过细胞传代 ,就获得了能稳定高效表达 EGFP的绿色细胞 .比较发现 ,质粒 p CA13 - e G转染后 ,产生能高效表达 EGFP的 He L a细胞其比率高于 Vero细胞 ;EGFP高效表达对 Vero细胞的毒性大于对 He L a细胞的毒性 .本研究表明 ,绿色细胞轮廓清晰 ,由于其特有的性质 ,在用于细胞的形态观察、细胞分裂等研究时会有所作为 .
p53 gene is a tumor suppressor gene and is involved in cell growth control.Wild type p53 gene was inserted into pCA13 and got recombinant plasmid pCAp53.293 cells(a kind of adenovirus packing cells)was co transfected with pCAp53 and pBHG11,recombinant p53 adenovirus was obtained.Virus particle can be seen under electric microscopy and its diameter is about 80 nm.Virus titer is 6.7×10 8 PFU/mL.Recombinant p53 adenovirus have no influence on growth of normal lung cells,but it can cause death of bladder tumor cell (EJ)and primary culture breast cancer cell.MTT assay results showed that recombinant p53 adenovirus inhibited growth of p53 deficient tumor cell,such as bladder tumor cell(EJ)and epidermis cancer cell(A431)and inhibiting rate in 72 hours may be 86.7% and 78.4%.Flow cytometry test showed that recombinant p53 adenovirus induced apoptosis of tumor cell.Wild type p53 gene mediated by adenovirus vector may be used in the treatment for cancer.
