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目的 :克隆和在 COS- 7细胞中表达人血管内皮生长因子 16 5 (VEGF16 5 )。方法 :利用 RT- PCR方法 ,从新鲜卵巢癌组织中扩增得到人 VEGF16 5的全长 c DNA,并测定其核酸序列 ;利用基因重组技术构建 VEGF16 5的真核表达载体 pc DNA3.1- VEGF16 5 ;应用脂质体介导的基因转移技术将这一表达载体导入 COS- 7细胞后 ,免疫组化 (SP法 )检测瞬时表达的产物。结果 :经酶切鉴定和基因测序证实 ,克隆的基因片段为人 VEGF16 5 c DNA,重组质粒 pc DNA3.1- VEGF16 5转染 COS- 7细胞后 ,免疫组化检测有 VEGF表达。结论 :成功克隆人 VEGF 16 5基因 ,并在 COS- 7细胞中获得表达。
AIM: To clone and express human vascular endothelial growth factor 165 (VEGF165) in COS-7 cells. Methods: The full-length c DNA of human VEGF16 5 was amplified by RT-PCR from fresh ovarian cancer tissues and its nucleic acid sequence was determined. The eukaryotic expression vector pcDNA3.1-VEGF16 of VEGF16 5 was constructed by gene recombination technology 5; using liposome-mediated gene transfer technology to introduce this expression vector into COS-7 cells, immunohistochemistry (SP method) to detect the transient expression of the product. Results: The recombinant plasmid pcDNA3.1-VEGF165 was transfected into COS-7 cells by restriction endonuclease digestion and sequencing. The VEGF gene expression was detected by immunohistochemistry. Conclusion: The VEGF165 gene was successfully cloned and expressed in COS-7 cells.